JP2011012045A - Undifferentiation maintenance promoting agent for melanocyte stem cell - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an agent for suppressing depigmentation and aging of hair.SOLUTION: The depigmentation and aging of hair are suppressed with one or more proteins included in a protein group activated by a genome damage inducing substance.

Description

The present invention relates to a pigment stem cell undifferentiation maintenance promoter, self-renewal maintenance promoter, tissue stem cell undifferentiation maintenance promoter, mammalian depigmentation therapeutic agent, anti-aging therapeutic agent, cosmetic composition, and functionality. Regarding food.

In general, with aging, various aging phenomena such as a decrease in motor function and visceral function, a decrease in immune function, a decrease in memory ability, skin spots, wrinkles, presbyopia, and gray hair appear. The present inventor focuses on gray hair, which is easy to follow, and studies the cause of the aging phenomenon.

Hair plays an important role in identifying and defending individuals by color and pattern.
Hair is formed in hair follicles including hair papilla and hair matrix cells, and pigment cells exist between the hair matrix cells. When pigment cells differentiate in the skin, they produce melanin pigments and pass them to the surrounding keratinocytes to supply pigments to the skin and hair. And so on. It was known that the number of pigment cells in hair follicles that grow white hair was reduced, but the mechanism was hardly known.

The present inventors have found pigment stem cells so far (Non-patent Document 1) and have clarified that gray hair is caused by disappearance of pigment stem cells (Non-patent Document 2).

Nishimura EK et al. Nature. 416 (6883): 854-60, 2002 Nishimura EK et al. Science. 307 (5710): 720-724. 2005

As described above, it is clear that gray hair is caused by disappearance of pigment stem cells, but what mechanism is causing depigmentation (whitening) of hair with aging, that is, aging It has not yet been clarified how the pigment stem cells have disappeared. The elucidation of this mechanism is desired in the field of development of pharmaceuticals and bio-related products aiming at clarifying the cause of the aging phenomenon and controlling aging and suppressing depigmentation of hair.

In particular, various methods such as administration of a vasodilator and a plant extract have been taken for the purpose of suppressing depigmentation of hair. Therefore, development of a depigmentation inhibitor for hair is desired.

The present invention has been made in view of the above circumstances, and provides a pharmaceutical for clarifying the cause of depigmentation of hair with aging and the cause of aging phenomenon, and suppressing depigmentation and aging of hair. With the goal.

According to the present invention, Ataxia telangiectasia mutated (A
TM) or a pigment stem cell undifferentiated maintenance promoting agent comprising an ATM variant having an action to promote undifferentiated maintenance of pigment cells is provided. The present invention maintains and promotes the undifferentiation of pigment stem cells with ATM or ATM variants, as shown in the Examples below.

Further, according to the present invention, a promoter sequence for controlling ATM or a variant thereof to be expressed in a region containing a pigment stem cell, and a nucleotide encoding an ATM or ATM variant linked downstream of the promoter sequence. An agent for maintaining undifferentiation of pigment stem cells, comprising a vector having the sequence, is provided. The present invention maintains and promotes the undifferentiation of pigment stem cells with ATM or ATM variants, as shown in the Examples below.

In addition, according to the present invention, there is provided an agent for maintaining undifferentiation of pigment stem cells, comprising ATM and Rad3-related (ATR) or an ATR variant having an effect of promoting the maintenance of pigment cell undifferentiation. It will be apparent to those skilled in the art that the present invention maintains and promotes the undifferentiation of pigment stem cells with ATR or ATR mutants, as shown in the Examples below.

In addition, according to the present invention, a promoter sequence for controlling ATR or a variant thereof to be expressed in a region containing pigment stem cells, and a nucleotide encoding ATR or an ATR variant linked downstream of the promoter sequence. An agent for maintaining undifferentiation of pigment stem cells, comprising a vector having the sequence, is provided. It will be apparent to those skilled in the art that the present invention maintains and promotes the undifferentiation of pigment stem cells with ATR or ATR mutants, as shown in the Examples below.

In addition, according to the present invention, there is provided an agent for maintaining undifferentiation of pigment stem cells, which comprises one or more proteins included in a protein group activated by a genome damage inducer. The present invention maintains and promotes the undifferentiation of pigment stem cells with one or more proteins included in a protein group activated by a genome damage inducer, as shown in the Examples below.

In addition, according to the present invention, a promoter sequence that is controlled to be expressed in a region including a pigment stem cell, and a kind of protein included in a group of proteins activated by a genome damage inducer that is connected downstream of the promoter sequence. An agent for maintaining the undifferentiation of pigment stem cells, comprising a vector having a nucleotide sequence encoding the above protein, is provided. The present invention maintains and promotes the undifferentiation of pigment stem cells with one or more proteins included in a protein group activated by a genome damage inducer, as shown in the Examples below.

In addition, according to the present invention, there is provided an agent for maintaining the undifferentiation of pigment stem cells, which comprises a regulatory factor that controls the activity of one or more proteins contained in a protein group activated by a genome damage-inducing source. The present invention maintains and promotes the undifferentiation of pigment stem cells by a control factor that controls the activity of one or more proteins contained in a protein group activated by a genome damage-inducing agent, as shown in the Examples below. .

Further, according to the present invention, ATM or ATM having an action of promoting self-replication maintenance of pigment cells.
An agent for maintaining the self-renewal of a pigment stem cell, comprising a mutant, is provided. The present invention promotes maintenance of the ability of pigment stem cells to self-replicate by ATM or ATM variants, as shown in the Examples below.

Further, according to the present invention, a promoter sequence for controlling ATM or a variant thereof to be expressed in a region containing pigment stem cells, and a nucleotide encoding an ATM or ATM variant linked downstream of the promoter sequence. And a self-replication promoting agent for pigment stem cells, comprising a vector having the sequence. The present invention promotes maintenance of the ability of pigment stem cells to self-replicate by ATM or ATM variants, as shown in the Examples below.

In addition, according to the present invention, ATR or ATR having an action of promoting the self-renewal of pigment cells.
An agent for maintaining the self-renewal of a pigment stem cell, comprising a mutant, is provided. It will be apparent to those skilled in the art that the present invention promotes the maintenance of the ability of pigment stem cells to self-replicate by ATR or ATR variants, as shown in the Examples below.

In addition, according to the present invention, a promoter sequence that controls ATR or a variant thereof to be expressed in a region containing pigment stem cells, and a nucleotide encoding ATR or an ATR variant that is linked downstream of the promoter sequence. And a self-replication promoting agent for pigment stem cells, comprising a vector having the sequence. It will be apparent to those skilled in the art that the present invention promotes the maintenance of the ability of pigment stem cells to self-replicate by ATR or ATR variants, as shown in the Examples below.

In addition, according to the present invention, there is provided an agent for maintaining the self-replication of pigment stem cells, which comprises one or more proteins included in a protein group activated by a genome damage inducer. The present invention promotes the maintenance of the ability of pigment stem cells to self-replicate with one or more proteins contained in a group of proteins activated by a genome damage inducer, as shown in the Examples below.

In addition, according to the present invention, a promoter sequence that is controlled to be expressed in a region including a pigment stem cell, and a kind of protein included in a group of proteins activated by a genome damage inducer that is connected downstream of the promoter sequence. An agent for promoting the maintenance of self-renewal of pigment stem cells, comprising a vector having a nucleotide sequence encoding the above protein, is provided. The present invention promotes the maintenance of the ability of pigment stem cells to self-replicate with one or more proteins contained in a group of proteins activated by a genome damage inducer, as shown in the Examples below.

In addition, according to the present invention, there is provided a pigment stem cell self-renewal maintenance promoter comprising a regulatory factor that controls the activity of one or more proteins contained in a protein group activated by a genome damage inducer. The present invention promotes the maintenance of the ability of pigment stem cells to self-replicate by a control factor that controls the activity of one or more proteins contained in a protein group activated by a genome damage inducer, as shown in the Examples below. To do.

In addition, according to the present invention, there is provided a therapeutic agent for depigmentation of a mammal, which contains the above-described agent for promoting maintenance of undifferentiation of pigment stem cells. As shown in the Examples described later, the present invention suppresses depigmentation by suppressing the depletion due to differentiation of the pigment stem cells by containing an agent that maintains and promotes the undifferentiation of pigment stem cells.

Moreover, according to this invention, the depigmentation therapeutic agent of mammal containing the self-replication inhibition inhibitor of the pigment stem cell mentioned above is provided. As shown in the examples below, the present invention includes an agent that promotes maintenance of the self-renewal ability of dye stem cells, thereby suppressing depletion due to loss of the self-replication ability of dye stem cells and suppressing depigmentation. .

Moreover, according to this invention, the undifferentiation maintenance promoter of the tissue stem cell containing the 1 or more types of protein contained in the protein group activated by the genome damage induction origin is provided. The present invention maintains and promotes the undifferentiation of tissue stem cells with one or more proteins included in a protein group activated by a genome damage inducer, as shown in the Examples below.

In addition, according to the present invention, a promoter sequence that is controlled to be expressed in a region containing tissue stem cells, and a kind of protein that is linked downstream of the promoter sequence and that is activated by a genome damage inducer An agent for maintaining undifferentiation of tissue stem cells, comprising a vector having a nucleotide sequence encoding the above protein, is provided. The present invention maintains and promotes the undifferentiation of tissue stem cells with one or more proteins included in a protein group activated by a genome damage inducer, as shown in the Examples below.

In addition, according to the present invention, there is provided an agent for maintaining undifferentiation of tissue stem cells, comprising a regulatory factor that controls the activity of one or more proteins contained in a protein group activated by a genome damage inducing agent. The present invention maintains and promotes the undifferentiation of tissue stem cells by a control factor that controls the activity of one or more proteins contained in a protein group activated by a genome damage inducer, as shown in the Examples below. .

In addition, according to the present invention, there is provided a therapeutic agent for suppressing aging in mammals, which comprises the above-described agent for promoting maintenance of undifferentiation of tissue stem cells. As shown in the Examples described later, the present invention suppresses depletion due to differentiation of tissue stem cells and suppresses aging by including an agent that maintains and promotes the undifferentiation of tissue stem cells.

Moreover, according to this invention, the cosmetic composition containing the 1 or more types of protein contained in the protein group activated by the genome damage induction origin is provided. In the present invention, as shown in Examples described later, one or more proteins included in a protein group activated by a genome damage inducer maintain and promote undifferentiation of tissue stem cells, thereby allowing differentiation of tissue stem cells. Suppresses the depletion caused by hair and maintains the youthful hair or bare skin.

Moreover, according to this invention, the functional food containing the 1 or more types of protein contained in the protein group activated by the genome damage induction origin is provided. The present invention, as shown in the examples below,
One or more proteins included in the protein group activated by the genomic damage-inducing source maintain the youthful hair, bare skin, body environment, etc. by maintaining and promoting the undifferentiation of tissue stem cells.

According to the present invention, as shown in the examples below, the protein group activated by the genomic damage inducer promotes the maintenance of undifferentiation of pigment stem cells, promotes the maintenance of self-replicating ability, Maintains and promotes undifferentiation of stem cells and suppresses depigmentation and aging of mammals.

It is a figure which shows the body color and histological change of a mouse | mouth before and after irradiation. (A) is a whole body photograph of a mouse before irradiation. (B) Whole body photograph after irradiation (5 Gy IR (+)). Gray hair is induced only after irradiation. (C)-(j) show the histological change of the hair follicle after irradiation in the growth period V stage. (C) and (g) are hole mount images. (D), (h) are skin sections after lacZ staining of Dct-lacZ transgenic mice. In the bulge region of the control mouse, pigment stem cells having no melanin pigment can be confirmed (arrows in c and d). (E), (f), (i), (j) show lacZ positive cells in the bulge region. These cells express Kit at the same time, and do not have melanin when there is no irradiation in bright field observation (arrows e and f). When irradiated with radiation, it contains a brown to black pigment (arrowheads i and j). On the 7th day after irradiation with 5 Gy, normal pigment stem cells have small round vesicles, but are observed to have an ectopically colored pigment cell dendritic morphology (i, j Arrowhead). (K) to (n) show lacZ positive cells after irradiation. The lacZ positive cells disappear from the bulge area in the second hair cycle. (K) and (l) are hole mount images. (K), (m) are lacZ-stained skin of mice before irradiation. (L), (n) are lacZ stained skin of mice after irradiation. The bulge area (bg) is indicated by parentheses. (O) shows the ratio of lacZ positive cells per total follicle in each hair cycle of the first, second and third after irradiation. (P)-(s) show the histochemical changes in the bulge area after administration of a genomic damaging agent. (Q) is when busulfan (40 mg / kg) is administered. (R) is when mitomycin C (4 mg / kg body weight) is administered. (S) is when hydrogen peroxide (1% in PBS) is administered. On the 6th day after administration of each drug, ectopically colored melanocytes were confirmed (q to s) but not in the control (p). Gray hair was seen in the second hair cycle after administration of each drug (u to w), but not in the control (t). (X) shows a whole-mount lacZ-stained image in the bulge region of the eyelash accompanying the aging of Dct-lacZ mice. Dendritic pigment cells are found in the bulge region of the hair follicles of aging mice (arrowhead, 22 months old). (Y), (z) shows the bulge region of 42-week-old XPD ^{R722W / R722W} mice and XPD + / + mice. ^{PDR722W / R722W} mice have pigments in Kit + positive cells (z) in the hair follicle bulge (arrowheads) but are not confirmed in control mice (XPD + / + arrowheads of mouse mice) (y). It is a figure which shows the immunohistochemical analysis of (gamma) -H2AX and 53BP1 expression in a basil area | region. Seven-week-old mice in which almost all hair follicles were synchronized in the resting period were irradiated with radiation, and tissues were collected 6 hours after the irradiation. (A), (c) shows the dormant hair follicle which is not irradiated with radiation. (B), (d) shows a resting hair follicle 6 hours after irradiation. (E) and (g) show hair follicles not irradiated with radiation one day after hair removal. (F) and (h) show hair follicles 6 hours after radiation irradiation one day after hair removal. There was no difference in the distribution of genomic damage focus due to irradiation in the bulge region including Kit positive cells regardless of the presence or absence of hair removal (arrows b, f, d, h). (A) shows changes in the body color of ATM-deficient mice and control mice after irradiation with 3 Gy. The upper row shows the body color before irradiation, the middle row shows the first hair cycle after irradiation, and the lower row shows the body color in the second hair cycle after irradiation. In general, only 3My mice show clear whitening when irradiated with 3 Gy radiation that cannot induce whitening. (B) to (g) show immunofluorescence staining for detecting γ-H2AX damage focus formation in the bulge region after irradiation with 3 Gy. In ATM-deficient mice, the focus formation possessed by Kit-positive cells is retained even after 12 hours (g). The inset shows a magnified image of the cell containing the focus of γ-H2AX indicated by the white arrowhead. (H)-(j) show Dct-lacZ positive cells in the bulge region on day 5 after irradiation with 3 Gy in ATM-deficient mice and control mice. In ATM-deficient mice, even in the case of irradiation with 3 Gy, dendritic pigmented photoreceptor cells can be confirmed in the bulge region (j). The inset is an enlarged image of lacZ positive cells in the bulge region. The parentheses indicate the bulge area (bg). (K) shows the frequency of hair follicles having ectopically colored melanocytes in the bulge region for all hair follicles after irradiation with 3 Gy.

(Background of the invention)
The inventors of the present invention have studied the mechanism of gray hair with aging, and have ectopic pigment granules with dendritic structures prior to the occurrence of gray hair with aging. It was discovered that pigment cells appear in the bulge region (Non-patent Document 2). That is, in the bulge area
It was suggested that pigment stem cells were differentiated prior to the expression of gray hair. However, it was unclear whether this was equivalent to normal differentiation.

On the other hand, in young mice, undifferentiated pigment stem cells are distributed in the bulge region, and mature pigment cells are not detected. For this reason, pigment cells appearing ectopically in the bulge region are considered to be a phenomenon specifically related to the causes of aging and gray hair.

Moreover, early depigmentation is known to occur due to various genomic damage-inducing sources such as exposure to radiation that causes double-strand breaks in DNA as well as in progeria. In addition, Ataxia telangiectasia that serves as a monitoring mechanism for genomic damage
It is known that when the mutated (ATM) gene or the like does not function, the genome damage response is not properly performed, causing progeria.

Therefore, the present inventors focused on the possibility that the appearance of ectopic pigment cells in the bulge region may be involved in the genomic damage response reaction, and investigated the effect of irradiation on living mice. As a result, we discovered that pigment cells with ectopic pigment granules in the form of dendrites were expressed in place of undifferentiated pigment stem cells in the basil region in the middle growth phase after irradiation. These cells were also morphologically similar to ectopic differentiated pigment cells that appeared with aging. Therefore, when ectopic differentiated pigment cells induced after irradiation were analyzed by immunohistochemical staining and electron microscopy, it was revealed that the morphology and molecular expression were close to normal differentiation. Therefore, we conclude that the fate of pigment stem cells for aging and eliciting more than a certain genomic damage response is differentiation.

Furthermore, in the pigment cells differentiated ectopically in these bulge regions, the remaining of γ-H2AX (phosphorylated histone H2AX), which is a genome damage response marker, was observed. It was discovered that ectopicly differentiated pigment cells in the bulge region were eventually excluded from the bulge region in the late growth phase.

In addition, the present inventors also examined the effects of radiation on mice lacking the ATM gene, which is a major mediator of the genomic damage response. In general, in the genome damage response, ATM,
ATM and Rad3-related (ATR), H2AX (γ-H2AX), 5
It is known that various proteins such as 3BP1 are activated, and these protein groups cooperate to repair damaged DNA (J. Wada Harper et al. Molecular Cell. 28: 739-745. 2
007, Tom Misteli et al. Molecular Cell Biology. 10: 243-254. 2009). Therefore, by using an ATM-deficient mouse which is a kind of these protein groups, it is possible to examine the possibility that ectopic differentiation of pigment stem cells is performed as part of the genomic damage response. As a result, it was found that pigment stem cells lacking ATM cause differentiation into pigment cells more sensitively and cause earlier depigmentation compared to wild type.

From the above results, due to the failure of the genomic damage response, the pigment stem cells differentiate into ectopic pigment cells that take a dendritic form in the bulge region, and the differentiated pigment cells are excluded from the bulge region. It became clear that depletion occurred and whitening occurred.

In addition, in the analysis results of ATM-deficient mice, the protein group activated by the genome damage inducing agent such as ATM has a level where DNA damage can be repaired by activating the genome damage response pathway. , Suggesting that pigment stem cells contribute to the maintenance of stem cells by protecting them from fate decisions on differentiation.

Based on the above, one or more proteins included in the group of proteins activated by genomic damage-inducing agents are essential for maintaining the ability of pigment stem cells to self-replicate and undifferentiated. It became clear to cause. Therefore, one or more proteins included in the protein group activated by the genomic damage-inducing source can maintain and promote the self-renewal ability and undifferentiation of pigment stem cells, and depigmentation by maintaining pigment stem cells It became clear that it was possible to suppress this, and the present invention was completed.

In addition, the mechanism described above is not limited to pigment stem cells, and one or more proteins included in the protein group activated by the genomic damage-inducing source may be used to maintain the ability of tissue stem cells to self-renew and maintain undifferentiation. It is clear that it is important.

In addition, tissue stem cell depletion due to the accumulation of genomic damage has been experimentally shown in recent years in animals with genomic instability. It has been suggested to cause. This tissue stem cell depletion is considered to be caused by the same mechanism as the above-described dye stem cell depletion. Therefore, one or more proteins included in the protein group activated by the genomic damage-inducing source suppresses tissue stem cell depletion and suppresses aging by maintaining and promoting the self-replication ability and undifferentiation of tissue stem cells. It is considered possible to do.

<Explanation of terms>
In the present specification and claims, the meaning of each term is defined as follows.

(I) Hair Hair refers to an appendage of the skin composed of a hair shaft and hair root mainly composed of keratin, and includes, for example, head hair and body hair.

(Ii) Depigmentation of hair Hair depigmentation means that hair that originally has melanin pigments approaches the white color of the hair as the supply of melanin pigments from pigment cells decreases. . This includes, for example, the change of hair to white hair and whitening of body hair.

(Iii) Stem cells Stem cells refer to undifferentiated cells that exhibit high self-sustaining ability (self-replicating ability) and can supply differentiated progeny cells. Examples include pigment stem cells that supply pigment cells as progeny cells, and hair follicle stem cells that have the ability to form bulge regions as undifferentiated keratinocytes and differentiate into skin appendages such as epidermis, sweat glands, and sebaceous glands. .

(Iv) Genomic damage-inducing sources Genomic damage-inducing sources include active oxygen generated in daily metabolism, ultraviolet rays in the environment and food, radiation, carcinogens, and other factors that can cause genomic damage response (DDR). It is out.

(V) Protein group activated by genome damage inducer The protein group activated by genome damage inducer is a protein group activated by genome damage, and these proteins are genomes. The genome is repaired by collaborating in a damage response system. Examples of such proteins include ATM and A.
TR, H2AX (γ-H2AX), 53BP1 and the like can be mentioned. In our somatic cells, about 10 to 1 million genome damages are generated per cell per day, and the genome damage response system repairs this genome damage, thereby keeping the genome stable.

(Vi) Regulatory factor A regulatory factor refers to a compound, oligonucleotide, peptide, protein or the like that controls the activity of a protein group activated by a genome damage inducer.

(Vii) Niche Niche refers to an ecological place for stem cells. Usually, the localized site of tissue stem cells is called a niche. In a niche environment, stem cells are maintained in an undifferentiated state. For example, the niche of pigment stem cells and hair follicle stem cells exists in the bulge region. Usually, some of the progeny cells grown from the pigment stem cells in the bulge region move from the bulge region to the hair matrix, where they differentiate and mature into pigment cells to supply the melanin pigment to the hair.

(Viii) Ectopic differentiation of pigmented stem cells Ectopic differentiation of pigmented stem cells refers to pigmented stem cells that are ectopically (out of place) in a bulge region (niche) that is originally maintained in an undifferentiated state in wild-type mice. Means that it has matured differentiation, such as having melanin granules and acquiring a dendritic morphology. Normally, this ectopic differentiated cell disappears without being maintained. For example, it means that a pigment stem cell loses undifferentiation in a bulge region and differentiates into a mature pigment cell.

(Ix) Amino acid sequence homology (%)
Amino acid sequence homology (%) refers to the state in which sequences are aligned and gaps are introduced as necessary to obtain maximum percent sequence identity, and any conservative substitutions are not considered part of sequence identity. Defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues of a particular amino acid sequence. The% identity value used here is WU-BLAST-2.
(Altschul et al., Methods in Enzymology 266: 46
0-480 (1996)). WU-BLAST-2 uses several search parameters, most of which are set to default values. The adjustable parameters are set to the following values: overlap span = 1, overlap fraction = 0.
125, word threshold (T) = 11. The% amino acid sequence identity value is determined by dividing the number of matched identical residues by the total number of residues in the aligned region.

(X) Base sequence homology (%)
Base sequence homology (%) refers to the state where sequences are aligned and gaps are introduced as necessary to obtain maximum percent sequence identity and any conservative substitutions are not considered part of the sequence identity. Defined as the percentage of nucleotide residues in a candidate sequence that are identical to nucleotide residues in a particular base sequence. Alignments for determining nucleotide sequence identity can be determined by various methods known to those skilled in the art, such as BLAST, BLAST-2, AL
This can be accomplished by using publicly available computer software such as IGN or Megalign (DNASTAR) software. One skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. For example, a particular fragment or subregion of two sequences is recognized to be more or less homologous than compared between all its own fragments.

(Xi) Deletion / Substitution / Addition Polypeptide variants can be in a different form from the native protein. A “substitution” variant is
At least one amino acid in the native sequence is removed and a different amino acid is inserted at the same position in the place. A substitution can be a single substitution in which only one amino acid in the molecule is substituted, or a multiple substitution in which two or more amino acids are substituted in the same molecule.
An “insert” variant is one in which one or more amino acids have been inserted immediately adjacent to the amino acid at a particular position in the native sequence. Immediately adjacent to an amino acid means bound to either the α-carboxyl or α-functional group of the amino acid. A “deletion” variant is one in which one or more amino acids in the native amino acid sequence have been removed. Usually, deletion mutants are
One or two amino acids are deleted in a particular region of the molecule. Peptide variants also include covalent modifications to the residues in addition to the epitope tag heterologous polypeptide.

(Xii) Stringency The “stringency” of a hybridization reaction can be readily determined by one skilled in the art and is generally calculated empirically depending on probe length, wash temperature, and salinity.
In general, longer probes require higher temperatures for proper annealing, while shorter probes require lower temperatures. Hybridization generally relies on the ability of denatured DNA to reanneal when complementary strands are present in an environment close to, but below, the Tm (melting temperature). The higher the degree of desired homology between the probe and hybridizable sequence, the higher the relative temperature that can be used. As a result, higher relative temperatures result in more stringent reaction conditions, while lower temperatures do not. Furthermore, stringency is inversely proportional to salinity. For further details and explanation of the stringency of hybridization reactions, see Au
subel et al., Protocol in Molecular Biology (19
95).

“Stringent conditions” include: (1) using low ionic strength and high temperature for washing, eg, 0.015M sodium chloride / 0.0015M sodium citrate / 0.1% sodium dodecyl sulfate at 50 ° C .; ) Denaturing agents such as formamide, for example 0.1% bovine serum albumin / 0.1% Ficoll / 0.1% polyvinylpyrrolidone / 750 mM sodium chloride, pH 6.5 50 mM sodium phosphate with 75 mM sodium citrate Illustrated by the reaction conditions characterized by using 50% (volume / volume) formamide with a buffer at 42 ° C. Alternatively, stringent conditions are 50% formamide, 5
xSSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5 × Denhart solution, sonicated sperm DNA (50 μg / ml), 0 1% SDS and 10% dextran sulfate at 42 ° C
And washed with 0.2 × SSC (sodium chloride / sodium citrate) and 50% formaldehyde at 55 ° C., followed by a high stringency wash consisting of 0.1 × SSCV containing EDTA at 55 ° C. One skilled in the art knows how to adjust temperature, ionic strength, etc. to adapt to factors such as probe length as needed.

Hereinafter, embodiments of the present invention will be described in detail.
<Embodiment 1: Undifferentiated maintenance promoter containing ATM or ATM variant>
The present embodiment is an agent for promoting the maintenance of undifferentiated pigment stem cells, which comprises ATM or an ATM variant having an effect of promoting the maintenance of undifferentiated pigment cells. As shown in the Examples below, the present inventors have also found mice lacking the ATM gene, which is a major mediator of the genomic damage response,
We are investigating the effects of radiation In general, for genomic damage response, ATM, ATR, H2A
It is known that various proteins such as X (γ-H2AX) and 53BP1 are activated, and these protein groups cooperate to repair damaged DNA (J. Wada Harper et al. Molecul
ar Cell. 28: 739-745. 2007, Tom Misteli et al. Molecular Cell Biology. 10: 243-254
2009). Therefore, by using an ATM-deficient mouse which is a kind of these protein groups, it is possible to examine the possibility that ectopic differentiation of pigment stem cells is performed as part of the genomic damage response. As a result, in pigment stem cells lacking ATM, compared to wild type,
It has been found that it causes differentiation into pigment cells more sensitively and causes earlier depigmentation. According to this experiment, it can be said that ATM or an ATM variant has a function of maintaining and promoting undifferentiation of pigment stem cells, and thus it can be said that the agent itself containing them also maintains and promotes undifferentiation of pigment stem cells.

As described above, the present inventors have found that the ectopic differentiation of the pigment stem cells described above is caused when the genome of the pigment stem cells is damaged by the genome damage inducer. That is,
Promoting the maintenance of undifferentiated pigment stem cells refers to suppressing the differentiation of pigment stem cells into mature pigment cells in the bulge region, which is caused by such genomic damage-inducing sources. In addition,
The explanation of the promotion of undifferentiated maintenance of these pigment stem cells is common to other embodiments.

Here, when the agent of the present embodiment is used as, for example, a pharmaceutical, a cosmetic, or a functional food, there is a possibility that the effect can be obtained by oral administration or transdermal administration. Pills, tablets, capsules, creams, liquids for internal use, injections, and may be added to hair tonics, hair lotions, hair creams, aerosols, ointments, shampoos, rinses, etc. .

However, the specific use of the agent containing such ATM or ATM variant is not particularly limited to pharmaceutical use. For example, in an artificial organ / artificial tissue culture bioreactor used for regenerative medicine, ATM or an ATM variant is added to an appropriate concentration to supply artificial organs / artificial tissues used for regenerative medicine It can also be used to maintain the undifferentiated nature of stem cells as a source. That is, the specific use of an agent containing ATM or an ATM variant is not limited to in vivo use in mammals including humans, but in test tubes, bioreactors, artificial organs / artificial tissues. It is also possible to use the mammal in vitro. For example, in order to continuously cultivate artificially cultured hair in a healthy state, which is used for hair transplantation in the field of regenerative medicine, so as not to become gray hair, an appropriate concentration of ATM or ATM variant is obtained. Thus, it can also be used to maintain the undifferentiated nature of stem cells that are the source of artificial hair used in regenerative medicine. In addition, the description about these agents is common also to other embodiment.

<Embodiment 2: An undifferentiated maintenance promoter containing an ATM or ATM variant expression vector>
In the present embodiment, there is provided a promoter sequence that regulates ATM or a variant thereof to be expressed in a region containing pigment stem cells, and an ATM linked downstream of the promoter sequence.
Alternatively, an agent for maintaining undifferentiation of pigment stem cells, comprising a vector having a nucleotide sequence encoding an ATM variant. As shown in the examples described later, since an ATM or ATM variant has a function of maintaining and promoting undifferentiation of pigment stem cells by experiments using mice lacking the ATM gene, a nucleotide sequence encoding them The agent itself containing the vector having the above also promotes the maintenance of undifferentiation of pigment stem cells.

The term “promoter” as used herein refers to a sequence that controls the expression of a polypeptide present downstream of the promoter sequence, and in this case, a promoter that can be expressed in a region containing a pigment stem cell is particularly desirable. For example, K14 promoter, K5 promoter and the like can be mentioned. In addition, description of these promoters is common also to other embodiment.

The vector herein can be any vector that can be conveniently subjected to recombinant DNA procedures, and the choice of vector often depends on the host cell into which the vector is to be introduced. Thus, the vector may be a self-replicating vector, ie a vector that exists as extrachromosomal material whose replication is independent of chromosomal replication. Such vectors include, for example, a plasmid, phage, cosmid, mini, chromosome, or virus. Alternatively, the vector may be one that, when introduced into a host cell, is integrated into the host cell genome and replicated with one or more chromosomes integrated therein. Examples of suitable vectors are bacterial expression vectors and yeast expression vectors. Examples of the viral vectors include adenovirus, retrovirus, SV40
Examples include vectors derived from viruses such as papovavirus, vaccinia virus, fowlpox virus, and pseudorabies virus, but are not limited thereto. With the expression vector constructed in the present invention, one or more proteins contained in a protein group activated by a genome damage inducer can be expressed at any place. In addition, description of these vectors is common also to other embodiment.

<Embodiment 3: Self-replication maintenance promoter containing ATM or ATM variant>
The present embodiment is an agent for promoting the maintenance of self-replication of pigment stem cells, which comprises ATM or an ATM variant having a pigment cell undifferentiation maintenance promoting effect. As shown in the Examples below, the present inventors have also found mice lacking the ATM gene, which is a major mediator of the genomic damage response,
We are investigating the effects of radiation In general, for genomic damage response, ATM, ATR, H2A
It is known that various proteins such as X (γ-H2AX) and 53BP1 are activated, and these protein groups cooperate to repair damaged DNA (J. Wada Harper et al. Molecul
ar Cell. 28: 739-745. 2007, Tom Misteli et al. Molecular Cell Biology. 10: 243-254
2009). Therefore, by using an ATM-deficient mouse which is a kind of these protein groups, it is possible to examine the possibility that ectopic differentiation of pigment stem cells is performed as part of the genomic damage response. As a result, in pigment stem cells lacking ATM, compared to wild type,
It has been found that it causes differentiation into pigment cells more sensitively and causes earlier depigmentation. As a result of this experiment, it has been demonstrated that ATMs or ATM variants have the function of promoting the maintenance of the ability of pigment stem cells to self-replicate, so that the agents containing them themselves also promote the maintenance of the ability of pigment stem cells to self-replicate. I can say that.

As described above, the present inventors have found that the ectopic differentiation of the pigment stem cells described above is caused when the genome of the pigment stem cells is damaged by the genome damage inducer. That is, the promotion of pigment stem cell self-renewal refers to suppression of inhibition of pigment stem cell self-replication in the bulge region caused by abnormal maturation of pigment stem cells into mature pigment cells caused by genomic damage induction. . In addition, description about the self-replication maintenance promotion of these pigment stem cells is common to other embodiments. Other points are the same as those in the first embodiment, and are not repeated.

<Embodiment 4: Self-replication maintenance promoter containing ATM or ATM variant expression vector>
In the present embodiment, there is provided a promoter sequence that regulates ATM or a variant thereof to be expressed in a region containing pigment stem cells, and an ATM linked downstream of the promoter sequence.
Alternatively, it is an agent for maintaining the self-renewal of pigment stem cells, comprising a vector having a nucleotide sequence encoding an ATM variant. As shown in the Examples below, ATM or ATM variants have a function of promoting the maintenance of the ability of pigment stem cells to self-replicate. Therefore, the agent itself containing a vector having a nucleotide sequence encoding them is also an agent of pigment stem cells. Promote maintenance of self-replicating ability. Other points are the same as those in the third embodiment, and thus will not be repeated.

<Embodiments 1-4: Description of ATM and ATM variants>
In common with the above Embodiments 1-4, the amino acid sequence of human ATM protein is known,
The amino acid sequence shown in SEQ ID NO: 1 (NCBI Reference Sequence: NP_000042.3 ataxia
telangiectasia mutated isoform 1 [Homo sapiens]).

Sequence number 1: It describes from N terminal to C terminal (3056aa)
1 mslvlndlli ccrqlehdra terkkevekf krlirdpeti khldrhsdsk qgkylnwdav
61 frflqkyiqk eteclriakp nvsastqasr qkkmqeissl vkyfikcanr raprlkcqel
121 lnyimdtvkd ssngaiygad csnillkdil svrkywceis qqqwlelfsv yfrlylkpsq
181 dvhrvlvari ihavtkgccs qtdglnskfl dffskaiqca rqeksssgln hilaaltifl
241 ktlavnfrir vcelgdeilp tllyiwtqhr lndslkevii elfqlqiyih hpkgaktqek
301 gayestkwrs ilynlydllv neishigsrg kyssgfrnia vkenlielma dichqvfned
361 trsleisqsy tttqressdy svpckrkkie lgwevikdhl qksqndfdlv pwlqiatqli
421 skypaslpnc elspllmils qllpqqrhge rtpyvlrclt evalcqdkrs nlessqksdl
481 lklwnkiwci tfrgisseqi qaenfgllga iiqgslvevd refwklftgs acrpscpavc
541 cltlalttsi vpgtvkmgie qnmcevnrsf slkesimkwl lfyqlegdle nstevppilh
601 snfphlvlek ilvsltmknc kaamnffqsv pecehhqkdk eelsfsevee lflqttfdkm
661 dfltivrecg iekhqssigf svhqnlkesl drcllglseq llnnysseit nsetlvrcsr
721 llvgvlgcyc ymgviaeeea ykselfqkak slmqcagesi tlfknktnee frigslrnmm
781 qlctrclsnc tkkspnkias gfflrlltsk lmndiadick slasfikkpf drgevesmed
841 dtngnlmeve dqssmnlfnd ypdssvsdan epgesqstig ainplaeeyl skqdllfldm
901 lkflclcvtt aqtntvsfra adirrkllml idsstleptk slhlhmylml lkelpgeeyp
961 lpmedvlell kplsnvcsly rrdqdvckti lnhvlhvvkn lgqsnmdsen trdaqgqflt
1021 vigafwhltk erkyifsvrm alvnclktll eadpyskwai lnvmgkdfpv nevftqflad
1081 nhhqvrmlaa esinrlfqdt kgdssrllka lplklqqtaf enaylkaqeg mremshsaen
1141 petldeiynr ksvlltliav vlscspicek qalfalcksv kenglephlv kkvlekvset
1201 fgyrrledfm ashldylvle wlnlqdteyn lssfpfilln ytniedfyrs cykvliphlv
1261 irshfdevks ianqiqedwk slltdcfpki lvnilpyfay egtrdsgmaq qretatkvyd
1321 mlksenllgk qidhlfisnl peivvellmt lhepanssas qstdlcdfsg dldpapnpph
1381 fpshvikatf ayisnchktk lksileilsk spdsyqkill aiceqaaetn nvykkhrilk
1441 iyhlfvslll kdiksglgga wafvlrdviy tlihyinqrp scimdvslrs fslccdllsq
1501 vcqtavtyck dalenhlhvi vgtliplvye qvevqkqvld llkylvidnk dnenlyitik
1561 lldpfpdhvv fkdlritqqk ikysrgpfsl leeinhflsv svydalpltr leglkdlrrq
1621 lelhkdqmvd imrasqdnpq dgimvklvvn llqlskmain htgekevlea vgsclgevgp
1681 idfstiaiqh skdasytkal klfedkelqw tfimltylnn tlvedcvkvr saavtclkni
1741 latktghsfw eiykmttdpm laylqpfrts rkkflevprf dkenpfegld dinlwiplse
1801 nhdiwiktlt cafldsggtk ceilqllkpm cevktdfcqt vlpylihdil lqdtneswrn
1861 llsthvqgff tsclrhfsqt srsttpanld sesehffrcc ldkksqrtml avvdymrrqk
1921 rpssgtifnd afwldlnyle vakvaqscaa hftallyaei yadkksmddq ekrslafeeg
1981 sqsttissls ekskeetgis lqdllleiyr sigepdslyg cgggkmlqpi trlrtyehea
2041 mwgkalvtyd letaipsstr qagiiqalqn lglchilsvy lkgldyenkd wcpeleelhy
2101 qaawrnmqwd hctsvskeve gtsyheslyn alqslrdref stfyeslkya rvkeveemck
2161 rslesvysly ptlsrlqaig elesigelfs rsvthrqlse vyikwqkhsq llkdsdfsfq
2221 epimalrtvi leilmekemd nsqrecikdi ltkhlvelsi lartfkntql peraifqikq
2281 ynsvscgvse wqleeaqvfw akkeqslals ilkqmikkld ascaannpsl kltyteclrv
2341 cgnwlaetcl enpavimqty lekavevagn ydgessdelr ngkmkaflsl arfsdtqyqr
2401 ienymkssef enkqallkra keevgllreh kiqtnrytvk vqreleldel alralkedrk
2461 rflckaveny incllsgeeh dmwvfrlcsl wlensgvsev ngmmkrdgmk iptykflplm
2521 yqlaarmgtk mmgglgfhev lnnlisrism dhphhtlfii lalananrde fltkpevarr
2581 sritknvpkq ssqldedrte aanriictir srrpqmvrsv ealcdayiil anldatqwkt
2641 qrkginipad qpitklknle dvvvptmeik vdhtgeygnl vtiqsfkaef rlaggvnlpk
2701 iidcvgsdgk errqlvkgrd dlrqdavmqq vfqmcntllq rntetrkrkl tictykvvpl
2761 sqrsgvlewc tgtvpigefl vnnedgahkr yrpndfsafq cqkkmmevqk ksfeekyevf
2821 mdvcqnfqpv fryfcmekfl dpaiwfekrl aytrsvatss ivgyilglgd rhvqniline
2881 qsaelvhidl gvafeqgkil ptpetvpfrl trdivdgmgi tgvegvfrrc cektmevmrn
2941 sqetlltive vllydplfdw tmnplkalyl qqrpedetel hptlnaddqe ckrnlsdidq
3001 sfnkvaervl mrlqeklkgv eegtvlsvgg qvnlliqqai dpknlsrlfp gwkawv //

The base sequence of the human ATM gene is also known in the same manner, and the following SEQ ID NO: 2 (NCBI R
eference Sequence: NM_000051.3 Homo sapiens ataxia telangiectasia mutated (AT
M), transcript variant 1, mRNA).

SEQ ID NO: 2: described from the 5 ′ end to the 3 ′ end (13147 bp mRNA)
1 ccggagcccg agccgaaggg cgagccgcaa acgctaagtc gctggccatt ggtggacatg
61 gcgcaggcgc gtttgctccg acgggccgaa tgttttgggg cagtgttttg agcgcggaga
121 ccgcgtgata ctggatgcgc atgggcatac cgtgctctgc ggctgcttgg cgttgcttct
181 tcctccagaa gtgggcgctg ggcagtcacg cagggtttga accggaagcg ggagtaggta
241 gctgcgtggc taacggagaa aagaagccgt ggccgcggga ggaggcgaga ggagtcggga
301 tctgcgctgc agccaccgcc gcggttgata ctactttgac cttccgagtg cagtgacagt
361 gatgtgtgtt ctgaaattgt gaaccatgag tctagtactt aatgatctgc ttatctgctg
421 ccgtcaacta gaacatgata gagctacaga acgaaagaaa gaagttgaga aatttaagcg
481 cctgattcga gatcctgaaa caattaaaca tctagatcgg cattcagatt ccaaacaagg
541 aaaatatttg aattgggatg ctgtttttag atttttacag aaatatattc agaaagaaac
601 agaatgtctg agaatagcaa aaccaaatgt atcagcctca acacaagcct ccaggcagaa
661 aaagatgcag gaaatcagta gtttggtcaa atacttcatc aaatgtgcaa acagaagagc
721 acctaggcta aaatgtcaag aactcttaaa ttatatcatg gatacagtga aagattcatc
781 taatggtgct atttacggag ctgattgtag caacatacta ctcaaagaca ttctttctgt
841 gagaaaatac tggtgtgaaa tatctcagca acagtggtta gaattgttct ctgtgtactt
901 caggctctat ctgaaacctt cacaagatgt tcatagagtt ttagtggcta gaataattca
961 tgctgttacc aaaggatgct gttctcagac tgacggatta aattccaaat ttttggactt
1021 tttttccaag gctattcagt gtgcgagaca agaaaagagc tcttcaggtc taaatcatat
1081 cttagcagct cttactatct tcctcaagac tttggctgtc aactttcgaa ttcgagtgtg
1141 tgaattagga gatgaaattc ttcccacttt gctttatatt tggactcaac ataggcttaa
1201 tgattcttta aaagaagtca ttattgaatt atttcaactg caaatttata tccatcatcc
1261 gaaaggagcc aaaacccaag aaaaaggtgc ttatgaatca acaaaatgga gaagtatttt
1321 atacaactta tatgatctgc tagtgaatga gataagtcat ataggaagta gaggaaagta
1381 ttcttcagga tttcgtaata ttgccgtcaa agaaaatttg attgaattga tggcagatat
1441 ctgtcaccag gtttttaatg aagataccag atccttggag atttctcaat cttacactac
1501 tacacaaaga gaatctagtg attacagtgt cccttgcaaa aggaagaaaa tagaactagg
1561 ctgggaagta ataaaagatc accttcagaa gtcacagaat gattttgatc ttgtgccttg
1621 gctacagatt gcaacccaat taatatcaaa gtatcctgca agtttaccta actgtgagct
1681 gtctccatta ctgatgatac tatctcagct tctaccccaa cagcgacatg gggaacgtac
1741 accatatgtg ttacgatgcc ttacggaagt tgcattgtgt caagacaaga ggtcaaacct
1801 agaaagctca caaaagtcag atttattaaa actctggaat aaaatttggt gtattacctt
1861 tcgtggtata agttctgagc aaatacaagc tgaaaacttt ggcttacttg gagccataat
1921 tcagggtagt ttagttgagg ttgacagaga attctggaag ttatttactg ggtcagcctg
1981 cagaccttca tgtcctgcag tatgctgttt gactttggca ctgaccacca gtatagttcc
2041 aggaacggta aaaatgggaa tagagcaaaa tatgtgtgaa gtaaatagaa gcttttcttt
2101 aaaggaatca ataatgaaat ggctcttatt ctatcagtta gagggtgact tagaaaatag
2161 cacagaagtg cctccaattc ttcacagtaa ttttcctcat cttgtactgg agaaaattct
2221 tgtgagtctc actatgaaaa actgtaaagc tgcaatgaat tttttccaaa gcgtgccaga
2281 atgtgaacac caccaaaaag ataaagaaga actttcattc tcagaagtag aagaactatt
2341 tcttcagaca acttttgaca agatggactt tttaaccatt gtgagagaat gtggtataga
2401 aaagcaccag tccagtattg gcttctctgt ccaccagaat ctcaaggaat cactggatcg
2461 ctgtcttctg ggattatcag aacagcttct gaataattac tcatctgaga ttacaaattc
2521 agaaactctt gtccggtgtt cacgtctttt ggtgggtgtc cttggctgct actgttacat
2581 gggtgtaata gctgaagagg aagcatataa gtcagaatta ttccagaaag ccaagtctct
2641 aatgcaatgt gcaggagaaa gtatcactct gtttaaaaat aagacaaatg aggaattcag
2701 aattggttcc ttgagaaata tgatgcagct atgtacacgt tgcttgagca actgtaccaa
2761 gaagagtcca aataagattg catctggctt tttcctgcga ttgttaacat caaagctaat
2821 gaatgacatt gcagatattt gtaaaagttt agcatccttc atcaaaaagc catttgaccg
2881 tggagaagta gaatcaatgg aagatgatac taatggaaat ctaatggagg tggaggatca
2941 gtcatccatg aatctattta acgattaccc tgatagtagt gttagtgatg caaacgaacc
3001 tggagagagc caaagtacca taggtgccat taatccttta gctgaagaat atctgtcaaa
3061 gcaagatcta cttttcttag acatgctcaa gttcttgtgt ttgtgtgtaa ctactgctca
3121 gaccaatact gtgtccttta gggcagctga tattcggagg aaattgttaa tgttaattga
3181 ttctagcacg ctagaaccta ccaaatccct ccacctgcat atgtatctaa tgcttttaaa
3241 ggagcttcct ggagaagagt accccttgcc aatggaagat gttcttgaac ttctgaaacc
3301 actatccaat gtgtgttctt tgtatcgtcg tgaccaagat gtttgtaaaa ctattttaaa
3361 ccatgtcctt catgtagtga aaaacctagg tcaaagcaat atggactctg agaacacaag
3421 ggatgctcaa ggacagtttc ttacagtaat tggagcattt tggcatctaa caaaggagag
3481 gaaatatata ttctctgtaa gaatggccct agtaaattgc cttaaaactt tgcttgaggc
3541 tgatccttat tcaaaatggg ccattcttaa tgtaatggga aaagactttc ctgtaaatga
3601 agtatttaca caatttcttg ctgacaatca tcaccaagtt cgcatgttgg ctgcagagtc
3661 aatcaataga ttgttccagg acacgaaggg agattcttcc aggttactga aagcacttcc
3721 tttgaagctt cagcaaacag cttttgaaaa tgcatacttg aaagctcagg aaggaatgag
3781 agaaatgtcc catagtgctg agaaccctga aactttggat gaaatttata atagaaaatc
3841 tgttttactg acgttgatag ctgtggtttt atcctgtagc cctatctgcg aaaaacaggc
3901 tttgtttgcc ctgtgtaaat ctgtgaaaga gaatggatta gaacctcacc ttgtgaaaaa
3961 ggttttagag aaagtttctg aaacttttgg atatagacgt ttagaagact ttatggcatc
4021 tcatttagat tatctggttt tggaatggct aaatcttcaa gatactgaat acaacttatc
4081 ttcttttcct tttattttat taaactacac aaatattgag gatttctata gatcttgtta
4141 taaggttttg attccacatc tggtgattag aagtcatttt gatgaggtga agtccattgc
4201 taatcagatt caagaggact ggaaaagtct tctaacagac tgctttccaa agattcttgt
4261 aaatattctt ccttattttg cctatgaggg taccagagac agtgggatgg cacagcaaag
4321 agagactgct accaaggtct atgatatgct taaaagtgaa aacttattgg gaaaacagat
4381 tgatcactta ttcattagta atttaccaga gattgtggtg gagttattga tgacgttaca
4441 tgagccagca aattctagtg ccagtcagag cactgacctc tgtgactttt caggggattt
4501 ggatcctgct cctaatccac ctcattttcc atcgcatgtg attaaagcaa catttgccta
4561 tatcagcaat tgtcataaaa ccaagttaaa aagcatttta gaaattcttt ccaaaagccc
4621 tgattcctat cagaaaattc ttcttgccat atgtgagcaa gcagctgaaa caaataatgt
4681 ttataagaag cacagaattc ttaaaatata tcacctgttt gttagtttat tactgaaaga
4741 tataaaaagt ggcttaggag gagcttgggc ctttgttctt cgagacgtta tttatacttt
4801 gattcactat atcaaccaaa ggccttcttg tatcatggat gtgtcattac gtagcttctc
4861 cctttgttgt gacttattaa gtcaggtttg ccagacagcc gtgacttact gtaaggatgc
4921 tctagaaaac catcttcatg ttattgttgg tacacttata ccccttgtgt atgagcaggt
4981 ggaggttcag aaacaggtat tggacttgtt gaaatactta gtgatagata acaaggataa
5041 tgaaaacctc tatatcacga ttaagctttt agatcctttt cctgaccatg ttgtttttaa
5101 ggatttgcgt attactcagc aaaaaatcaa atacagtaga ggaccctttt cactcttgga
5161 ggaaattaac cattttctct cagtaagtgt ttatgatgca cttccattga caagacttga
5221 aggactaaag gatcttcgaa gacaactgga actacataaa gatcagatgg tggacattat
5281 gagagcttct caggataatc cgcaagatgg gattatggtg aaactagttg tcaatttgtt
5341 gcagttatcc aagatggcaa taaaccacac tggtgaaaaa gaagttctag aggctgttgg
5401 aagctgcttg ggagaagtgg gtcctataga tttctctacc atagctatac aacatagtaa
5461 agatgcatct tataccaagg cccttaagtt atttgaagat aaagaacttc agtggacctt
5521 cataatgctg acctacctga ataacacact ggtagaagat tgtgtcaaag ttcgatcagc
5581 agctgttacc tgtttgaaaa acattttagc cacaaagact ggacatagtt tctgggagat
5641 ttataagatg acaacagatc caatgctggc ctatctacag ccttttagaa catcaagaaa
5701 aaagttttta gaagtaccca gatttgacaa agaaaaccct tttgaaggcc tggatgatat
5761 aaatctgtgg attcctctaa gtgaaaatca tgacatttgg ataaagacac tgacttgtgc
5821 ttttttggac agtggaggca caaaatgtga aattcttcaa ttattaaagc caatgtgtga
5881 agtgaaaact gacttttgtc agactgtact tccatacttg attcatgata ttttactcca
5941 agatacaaat gaatcatgga gaaatctgct ttctacacat gttcagggat ttttcaccag
6001 ctgtcttcga cacttctcgc aaacgagccg atccacaacc cctgcaaact tggattcaga
6061 gtcagagcac tttttccgat gctgtttgga taaaaaatca caaagaacaa tgcttgctgt
6121 tgtggactac atgagaagac aaaagagacc ttcttcagga acaattttta atgatgcttt
6181 ctggctggat ttaaattatc tagaagttgc caaggtagct cagtcttgtg ctgctcactt
6241 tacagcttta ctctatgcag aaatctatgc agataagaaa agtatggatg atcaagagaa
6301 aagaagtctt gcatttgaag aaggaagcca gagtacaact atttctagct tgagtgaaaa
6361 aagtaaagaa gaaactggaa taagtttaca ggatcttctc ttagaaatct acagaagtat
6421 aggggagcca gatagtttgt atggctgtgg tggagggaag atgttacaac ccattactag
6481 actacgaaca tatgaacacg aagcaatgtg gggcaaagcc ctagtaacat atgacctcga
6541 aacagcaatc ccctcatcaa cacgccaggc aggaatcatt caggccttgc agaatttggg
6601 actctgccat attctttccg tctatttaaa aggattggat tatgaaaata aagactggtg
6661 tcctgaacta gaagaacttc attaccaagc agcatggagg aatatgcagt gggaccattg
6721 cacttccgtc agcaaagaag tagaaggaac cagttaccat gaatcattgt acaatgctct
6781 acaatctcta agagacagag aattctctac attttatgaa agtctcaaat atgccagagt
6841 aaaagaagtg gaagagatgt gtaagcgcag ccttgagtct gtgtattcgc tctatcccac
6901 acttagcagg ttgcaggcca ttggagagct ggaaagcatt ggggagcttt tctcaagatc
6961 agtcacacat agacaactct ctgaagtata tattaagtgg cagaaacact cccagcttct
7021 caaggacagt gattttagtt ttcaggagcc tatcatggct ctacgcacag tcattttgga
7081 gatcctgatg gaaaaggaaa tggacaactc acaaagagaa tgtattaagg acattctcac
7141 caaacacctt gtagaactct ctatactggc cagaactttc aagaacactc agctccctga
7201 aagggcaata tttcaaatta aacagtacaa ttcagttagc tgtggagtct ctgagtggca
7261 gctggaagaa gcacaagtat tctgggcaaa aaaggagcag agtcttgccc tgagtattct
7321 caagcaaatg atcaagaagt tggatgccag ctgtgcagcg aacaatccca gcctaaaact
7381 tacatacaca gaatgtctga gggtttgtgg caactggtta gcagaaacgt gcttagaaaa
7441 tcctgcggtc atcatgcaga cctatctaga aaaggcagta gaagttgctg gaaattatga
7501 tggagaaagt agtgatgagc taagaaatgg aaaaatgaag gcatttctct cattagcccg
7561 gttttcagat actcaatacc aaagaattga aaactacatg aaatcatcgg aatttgaaaa
7621 caagcaagct ctcctgaaaa gagccaaaga ggaagtaggt ctccttaggg aacataaaat
7681 tcagacaaac agatacacag taaaggttca gcgagagctg gagttggatg aattagccct
7741 gcgtgcactg aaagaggatc gtaaacgctt cttatgtaaa gcagttgaaa attatatcaa
7801 ctgcttatta agtggagaag aacatgatat gtgggtattc cgactttgtt ccctctggct
7861 tgaaaattct ggagtttctg aagtcaatgg catgatgaag agagacggaa tgaagattcc
7921 aacatataaa tttttgcctc ttatgtacca attggctgct agaatgggga ccaagatgat
7981 gggaggccta ggatttcatg aagtcctcaa taatctaatc tctagaattt caatggatca
8041 cccccatcac actttgttta ttatactggc cttagcaaat gcaaacagag atgaatttct
8101 gactaaacca gaggtagcca gaagaagcag aataactaaa aatgtgccta aacaaagctc
8161 tcagcttgat gaggatcgaa cagaggctgc aaatagaata atatgtacta tcagaagtag
8221 gagacctcag atggtcagaa gtgttgaggc actttgtgat gcttatatta tattagcaaa
8281 cttagatgcc actcagtgga agactcagag aaaaggcata aatattccag cagaccagcc
8341 aattactaaa cttaagaatt tagaagatgt tgttgtccct actatggaaa ttaaggtgga
8401 ccacacagga gaatatggaa atctggtgac tatacagtca tttaaagcag aatttcgctt
8461 agcaggaggt gtaaatttac caaaaataat agattgtgta ggttccgatg gcaaggagag
8521 gagacagctt gttaagggcc gtgatgacct gagacaagat gctgtcatgc aacaggtctt
8581 ccagatgtgt aatacattac tgcagagaaa cacggaaact aggaagagga aattaactat
8641 ctgtacttat aaggtggttc ccctctctca gcgaagtggt gttcttgaat ggtgcacagg
8701 aactgtcccc attggtgaat ttcttgttaa caatgaagat ggtgctcata aaagatacag
8761 gccaaatgat ttcagtgcct ttcagtgcca aaagaaaatg atggaggtgc aaaaaaagtc
8821 ttttgaagag aaatatgaag tcttcatgga tgtttgccaa aattttcaac cagttttccg
8881 ttacttctgc atggaaaaat tcttggatcc agctatttgg tttgagaagc gattggctta
8941 tacgcgcagt gtagctactt cttctattgt tggttacata cttggacttg gtgatagaca
9001 tgtacagaat atcttgataa atgagcagtc agcagaactt gtacatatag atctaggtgt
9061 tgcttttgaa cagggcaaaa tccttcctac tcctgagaca gttcctttta gactcaccag
9121 agatattgtg gatggcatgg gcattacggg tgttgaaggt gtcttcagaa gatgctgtga
9181 gaaaaccatg gaagtgatga gaaactctca ggaaactctg ttaaccattg tagaggtcct
9241 tctatatgat ccactctttg actggaccat gaatcctttg aaagctttgt atttacagca
9301 gaggccggaa gatgaaactg agcttcaccc tactctgaat gcagatgacc aagaatgcaa
9361 acgaaatctc agtgatattg accagagttt caacaaagta gctgaacgtg tcttaatgag
9421 actacaagag aaactgaaag gagtggaaga aggcactgtg ctcagtgttg gtggacaagt
9481 gaatttgctc atacagcagg ccatagaccc caaaaatctc agccgacttt tcccaggatg
9541 gaaagcttgg gtgtgatctt cagtatatga attacccttt cattcagcct ttagaaatta
9601 tattttagcc tttattttta acctgccaac atactttaag tagggattaa tatttaagtg
9661 aactattgtg ggtttttttg aatgttggtt ttaatacttg atttaatcac cactcaaaaa
9721 tgttttgatg gtcttaagga acatctctgc tttcactctt tagaaataat ggtcattcgg
9781 gctgggcgca gcggctcacg cctgtaatcc cagcactttg ggaggccgag gtgagcggat
9841 cacaaggtca ggagttcgag accagcctgg ccaagagacc agcctggcca gtatggtgaa
9901 accctgtctc tactaaaaat acaaaaatta gccgagcatg gtggcgggca cctgtaatcc
9961 cagctactcg agaggctgag gcaggagaat ctcttgaacc tgggaggtga aggttgctgt
10021 gggccaaaat catgccattg cactccagcc tgggtgacaa gagcgaaact ccatctcaaa
10081 aaaaaaaaaa aaaaaacaga aacgtatttg gatttttcct agtaagatca ctcagtgtta
10141 ctaaataatg aagttgttat ggagaacaaa tttcaaagac acagttagtg tagttactat
10201 ttttttaagt gtgtattaaa acttctcatt ctattctctt tatcttttaa gcccttctgt
10261 actgtccatg tatgttatct ttctgtgata acttcataga ttgccttcta gttcatgaat
10321 tctcttgtca gatgtatata atctctttta ccctatccat tgggcttctt ctttcagaaa
10381 ttgtttttca tttctaatta tgcatcattt ttcagatctc tgtttcttga tgtcattttt
10441 aatgtttttt taatgttttt tatgtcacta attattttaa atgtctgtac ttgatagaca
10501 ctgtaatagt tctattaaat ttagttcctg ctgtttatat ctgttgattt ttgtatttga
10561 taggctgttc atccagtttt gtctttttga aaagtgagtt tattttcagc aaggctttat
10621 ctatgggaat cttgagtgtc tgtttatgtc atattcccag ggctgttgct gcacacaagc
10681 ccattcttat tttaatttct tggctttagg gtttccatac ctgaagtgta gcataaatac
10741 tgataggaga tttcccaggc caaggcaaac acacttcctc ctcatctcct tgtgctagtg
10801 ggcagaatat ttgattgatg cctttttcac tgagagtata agcttccatg tgtcccacct
10861 ttatggcagg ggtggaagga ggtacattta attcccactg cctgcctttg gcaagccctg
10921 ggttctttgc tccccatata gatgtctaag ctaaaagccg tgggttaatg agactggcaa
10981 attgttccag gacagctaca gcatcagctc acatattcac ctctctggtt tttcattccc
11041 ctcatttttt tctgagacag agtcttgctc tgtcacccag gctggagtgc agtggcatga
11101 tctcagctca ctgaaacctc tgcctcctgg gttcaagcaa ttctcctgcc tcagcctccc
11161 gagtagctgg gactacaggc gtgtgccaac acgcccggct aattttttgt atttttatta
11221 gagacggagt ttcaccgtgt tagccaggat ggtctcgatc gcttgacctc gtgatccacc
11281 ctcctcggcc tcccaaagtg ctgggattac aggtgtgagc caccgcgccc ggcctcattc
11341 ccctcatttt tgaccgtaag gatttcccct ttcttgtaag ttctgctatg tatttaaaag
11401 aatgttttct acattttatc cagcatttct ctgtgttctg ttggaaggga agggcttagg
11461 tatctagttt gatacatagg tagaagtgga acatttctct gtcccccagc tgtcatcata
11521 taagataaac atcagataaa aagccacctg aaagtaaaac tactgactcg tgtattagtg
11581 agtataatct cttctccatc cttaggaaaa tgttcatccc agctgcggag attaacaaat
11641 gggtgattga gctttctcct cgtatttgga ccttgaaggt tatataaatt tttttcttat
11701 gaagagttgg catttctttt tattgccaat ggcaggcact cattcatatt tgatctcctc
11761 accttcccct cccctaaaac caatctccag aactttttgg actataaatt tcttggtttg
11821 acttctggag aactgttcag aatattactt tgcatttcaa attacaaact taccttggtg
11881 tatctttttc ttacaagctg cctaaatgaa tatttggtat atattggtag ttttattact
11941 atagtaaatc aaggaaatgc agtaaactta aaatgtcttt aagaaagccc tgaaatcttc
12001 atgggtgaaa ttagaaatta tcaactagat aatagtatag ataaatgaat ttgtagctaa
12061 ttcttgctag ttgttgcatc cagagagctt tgaataacat cattaatcta ctctttagcc
12121 ttgcatggta tgctatgagg ctcctgttct gttcaagtat tctaatcaat ggctttgaaa
12181 agtttatcaa atttacatac agatcacaag cctaggagaa ataactaatt cacagatgac
12241 agaattaaga ttataaaaga tttttttttt gtaattttag tagagacagg gttgccattg
12301 tattccagcc ttggcgacag agcaagactc tgcctcaaaa aaaaaaaaaa aaaggttttg
12361 gcaagctgga actctttctct caaatgacta agatagaaaa ctgccaagga caaatgagga
12421 gtagttagat tttgaaaata ttaatcatag aatagttgtt gtatgctaag tcactgaccc
12481 atattatgta cagcatttct gatctttact ttgcaagatt agtgatacta tcccaataca
12541 ctgctggaga aatcagaatt tggagaaata agttgtccaa ggcaagaaga tagtaaatta
12601 taagtacaag tgtaatatgg acagtatcta acttgaaaag atttcaggcg aaaagaatct
12661 ggggtttgcc agtcagttgc tcaaaaggtc aatgaaaacc aaatagtgaa gctatcagag
12721 aagctaataa attatagact gcttgaacag ttgtgtccag attaagggag ataatagctt
12781 tcccacccta ctttgtgcag gtcatacctc cccaaagtgt ttacctaatc agtaggttca
12841 caaactcttg gtcattatag tatatgccta aaatgtatgc acttaggaat gctaaaaatt
12901 taaatatggt ctaaagcaaa taaaagcaaa gaggaaaaac tttggacagc gtaaagacta
12961 gaatagtctt ttaaaaagaa agccagtata ttggtttgaa atatagagat gtgtcccaat
13021 ttcaagtatt ttaattgcac cttaatgaaa ttatctattt tctatagatt ttagtactat
13081 tgaatgtatt actttactgt tacctgaatt tattataaag tgtttttgaa taaataattc
13141 taaaagc //

(1) Specific Examples of Embodiments 1 and 3 (Polypeptides) Here, the above-mentioned ATM or ATM variant includes (a) ATM consisting of the amino acid sequence represented by SEQ ID NO: 1, (b) SEQ ID NO: 1. An ATM variant comprising an amino acid sequence having at least 80% homology with the amino acid sequence represented, and having an activity of promoting the maintenance of undifferentiation or self-renewal of pigment stem cells; (c) a nucleotide represented by SEQ ID NO: 2 ATM encoded by the sequence, or (d) a nucleic acid that hybridizes under stringent conditions to the nucleotide sequence represented by SEQ ID NO: 2 or its complementary strand, and promotes the maintenance of undifferentiation of pigment stem cells or self-replication Any of the ATM variants having a maintenance promoting effect is included.

Thus, if the amino acid sequence has at least 80% homology with the amino acid sequence represented by SEQ ID NO: 1, it can act to promote undifferentiation maintenance or self-renewal maintenance of pigment stem cells in the examples described later. It is possible for those skilled in the fields of basic medicine, pharmacology, biochemistry, etc. to maintain and promote the undifferentiation or self-renewal ability of pigment stem cells, as well as the ATM consisting of the amino acid sequence represented by SEQ ID NO: 1 that has been demonstrated. Is clear. The homology is preferably 80% or more, more preferably 85% or more, still more preferably 90% or more, and most preferably 95% or more. Thus, the higher the homology, the higher the possibility of maintaining and promoting the undifferentiated or self-replicating ability of the pigment stem cells, as in the ATM consisting of the amino acid sequence represented by SEQ ID NO: 1.

Alternatively, even in the case of an ATM variant having an amino acid sequence in which one or several amino acid residues are deleted, substituted, or added to the amino acid sequence represented by SEQ ID NO: 1, the pigment stem cell in the examples to be described later As in the case of ATM consisting of the amino acid sequence represented by SEQ ID NO: 1, which has been demonstrated to maintain and promote differentiation or self-replicating ability, it is essential to maintain and promote undifferentiated or self-replicating ability of pigment stem cells. It will be apparent to those skilled in the fields of medicine, pharmacy, biochemistry and the like. The term “several amino acid residues” as used herein means 2 to 9 amino acid residues, particularly preferably 2 to 5 amino acid residues, and more preferably 2 to 3 amino acid residues. Most preferred. Thus, the smaller the number of deletions / substitutions / additions of amino acid residues, the better the maintenance of undifferentiated or self-replicating ability of chromogenic stem cells, as in the ATM consisting of the amino acid sequence represented by SEQ ID NO: 1. Is more likely to do.

On the other hand, an AT having an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions to the nucleotide sequence represented by SEQ ID NO: 2 or its complementary strand as described above.
An AT encoded by the nucleotide sequence represented by SEQ ID NO: 2 that has been demonstrated to maintain and promote undifferentiated or self-replicating ability of pigment stem cells in the examples described later even for M mutants
Like M, maintaining and promoting undifferentiated or self-replicating ability of pigment stem cells is essential medicine,
It will be apparent to those skilled in the fields of pharmacy, biochemistry and the like. Here, stringent conditions can be easily determined by those skilled in the art. Generally, the probe length, washing temperature,
Although it is calculated empirically according to the salinity concentration, for example, the specific conditions described above in the explanation of terms can be suitably used.

In addition, as long as it is an ATM variant having an amino acid sequence encoded by a nucleotide sequence having at least 80% homology with the nucleotide sequence represented by SEQ ID NO: 2, the undifferentiated nature of the pigment stem cells in the examples described later Alternatively, as in the case of ATM encoded by the nucleotide sequence represented by SEQ ID NO: 2, which has been demonstrated to maintain and promote self-replicating ability, it is possible to maintain and promote undifferentiated or self-replicating ability of pigment stem cells. It will be apparent to those skilled in the fields of pharmacy, biochemistry and the like. The homology is preferably 80% or more, more preferably 85% or more, still more preferably 90% or more, and most preferably 95% or more. Thus, the higher the homology, the higher the possibility of maintaining and promoting the undifferentiation or self-replicating ability of the pigment stem cells, similar to the ATM encoded by the nucleotide sequence represented by SEQ ID NO: 2.

Furthermore, even in the case of an ATM variant encoded by a nucleotide sequence obtained by deleting, substituting, or adding one or several nucleotide residues to the nucleotide sequence represented by SEQ ID NO: 2, examples described later In the same manner as the ATM encoded by the nucleotide sequence represented by SEQ ID NO: 2, which has been demonstrated to maintain and promote undifferentiated or self-replicating ability of dye stem cells, the undifferentiated or self-replicating ability of dye stem cells is maintained and promoted. It will be apparent to those skilled in the fields of basic medicine, pharmacy, biochemistry and the like. In addition, although several nucleotide residues here mean 2 or more and 9 or less nucleotide residues, It is especially preferable that they are 2 or more and 5 or less nucleotide residues, and it is 2 or more and 3 or less nucleotide residues. Is most preferred. Thus, as the number of nucleotide residue deletions / substitutions / additions decreases, the undifferentiated or self-replicating ability of the pigment stem cell is maintained, as in the ATM encoded by the nucleotide sequence represented by SEQ ID NO: 2. The possibility to promote increases.

(1) Specific Examples of Embodiments 2 and 4 (Vector) On the other hand, the nucleotide sequence encoding the above ATM or ATM variant is (a) a nucleotide sequence encoding ATM consisting of the amino acid sequence represented by SEQ ID NO: 1. (B) A comprising an amino acid sequence having at least 80% homology with the amino acid sequence represented by SEQ ID NO: 1, and having an action of promoting the maintenance of undifferentiation of pigment stem cells or the maintenance of self-replication
A nucleotide sequence encoding a TM variant, (c) a nucleotide sequence represented by SEQ ID NO: 2, or (d) a nucleotide sequence represented by SEQ ID NO: 2 or a complementary strand thereof under stringent conditions, and a dye It is preferably any one of nucleotide sequences encoding an ATM variant having a stem cell undifferentiation maintenance promoting action or a self-renewal maintenance promoting action.

Thus, if it is a nucleotide sequence encoding ATM consisting of an amino acid sequence having at least 80% homology with the amino acid sequence represented by SEQ ID NO: 1, undifferentiated or self-replicating ability of pigment stem cells in the examples described later In the same manner as the nucleotide sequence encoding ATM consisting of the amino acid sequence represented by SEQ ID NO: 1 that has been demonstrated to maintain and promote the ATM, the ATM or ATM variant encoded by the nucleotide sequence is undifferentiated or self-differentiated in pigment stem cells. It is obvious to those skilled in the fields of basic medicine, pharmacy, biochemistry, and the like to maintain and promote the replication ability. The homology is preferably 80% or more, more preferably 85% or more, still more preferably 90% or more, and most preferably 95% or more. Thus, the higher the homology is, the ATM or ATM mutation that maintains and promotes the undifferentiated or self-replicating ability of pigment stem cells, as in the nucleotide sequence encoding ATM consisting of the amino acid sequence represented by SEQ ID NO: 1. The possibility of developing the body is increased.

Alternatively, even a nucleotide sequence encoding an ATM variant having an amino acid sequence in which one or several amino acid residues are deleted, substituted, or added to the amino acid sequence represented by SEQ ID NO: 1 In addition to the nucleotide sequence encoding ATM consisting of the amino acid sequence represented by SEQ ID NO: 1, which has been demonstrated to maintain and promote undifferentiated or self-replicating ability of pigment stem cells, ATM or ATM encoded by the nucleotide sequence It is essential that mutants maintain and promote the differentiation or self-renewal ability of pigment stem cells,
It will be apparent to those skilled in the field of biochemistry and the like. The term “several amino acid residues” as used herein means 2 to 9 amino acid residues, particularly preferably 2 to 5 amino acid residues, and more preferably 2 to 3 amino acid residues. Most preferred. in this way,
As the number of amino acid residue deletions / substitutions / additions decreases, the undifferentiated or self-replicating ability of chromogenic stem cells is maintained as in the nucleotide sequence encoding ATM consisting of the amino acid sequence represented by SEQ ID NO: 1. The possibility of expressing the ATM or ATM variant that promotes is increased.

On the other hand, even in the case of the nucleotide sequence represented by SEQ ID NO: 2 or a nucleotide sequence that hybridizes under stringent conditions as described above, undifferentiated or self-replicating ability of pigment stem cells in the examples described later In the same manner as the nucleotide sequence represented by SEQ ID NO: 2, which has been demonstrated to express ATM that maintains and promotes, it can express ATM or an ATM variant that maintains and promotes the undifferentiation or self-renewal ability of pigment stem cells. Basic medicine,
It will be apparent to those skilled in the fields of pharmacy, biochemistry and the like. Here, stringent conditions can be easily determined by those skilled in the art. Generally, the probe length, washing temperature,
The specific conditions described above can be suitably used, for example, depending on the salt concentration.

In addition, as long as the nucleotide sequence is at least 80% homologous to the nucleotide sequence represented by SEQ ID NO: 2, an ATM that maintains and promotes the undifferentiation or self-replicating ability of the pigment stem cells in the examples described later is used. Similar to the nucleotide sequence represented by SEQ ID NO: 2 that has been demonstrated to be expressed, ATM that maintains and promotes the undifferentiation or self-renewal ability of pigment stem cells
Alternatively, it will be apparent to those skilled in the fields of basic medicine, pharmacy, biochemistry, etc. that ATM variants are expressed. The homology is preferably 80% or more, more preferably 85% or more, still more preferably 90% or more, and most preferably 95%.
That's it. Thus, the higher the homology is, the higher the possibility of expressing ATM or an ATM variant that maintains and promotes the undifferentiation or self-renewal ability of pigment stem cells, as in the nucleotide sequence represented by SEQ ID NO: 2. Become.

Furthermore, in addition to the nucleotide sequence represented by SEQ ID NO: 2, even if it is a nucleotide sequence in which one or several nucleotide residues are deleted, substituted, or added, undifferentiated pigment stem cells in the examples described later Maintaining and promoting undifferentiated or self-replicating ability of chromogenic stem cells in the same manner as ATM encoded by the nucleotide sequence represented by SEQ ID NO: 2 which has been demonstrated to express ATM that maintains and promotes sex or self-renewal ability However, it will be apparent to those skilled in the fields of basic medicine, pharmacy, biochemistry and the like. In addition, although several nucleotide residues here mean 2 or more and 9 or less nucleotide residues, It is especially preferable that they are 2 or more and 5 or less nucleotide residues, and it is 2 or more and 3 or less nucleotide residues. Is most preferred. Thus, the smaller the number of nucleotide residue deletions / substitutions / additions, the ATM or the ability to maintain and promote the undifferentiation or self-renewal ability of pigment stem cells, as in the nucleotide sequence represented by SEQ ID NO: 2. The possibility of expressing ATM variants is increased.

<Embodiment 5: Undifferentiated maintenance promoter containing ATR or ATR mutant>
The present embodiment is an agent for promoting the maintenance of undifferentiated pigment stem cells, which comprises an ATR variant having an ATR or pigment cell undifferentiated maintenance promoting effect. As shown in the Examples below, the present inventors have also found mice lacking the ATM gene, which is a major mediator of the genomic damage response,
We are investigating the effects of radiation In general, for genomic damage response, ATM, ATR, H2A
It is known that various proteins such as X (γ-H2AX) and 53BP1 are activated, and these protein groups cooperate to repair damaged DNA (J. Wada Harper et al. Molecul
ar Cell. 28: 739-745. 2007, Tom Misteli et al. Molecular Cell Biology. 10: 243-254
2009). Therefore, by using an ATM-deficient mouse which is a kind of these protein groups, it is possible to examine the possibility that ectopic differentiation of pigment stem cells is performed as part of the genomic damage response. As a result, in pigment stem cells lacking ATM, compared to wild type,
It has been found that it causes differentiation into pigment cells more sensitively and causes earlier depigmentation.
Here, both ATM and ATR are members of the PI-3 kinase family of serine-threonine kinases and are structurally similar, and are located between the sensor and the effector in the genomic damage response, and both are central as transducers. (J. Wada Harper et al. Molecular Cell. 28: 739-745. 2007, Tom Miste
li et al. Molecular Cell Biology. 10: 243-254. 2009). In other words, it is well known that the roles of ATM and ATR partially overlap and work together (J. Wada Harper et al. M
olecular Cell. 28: 739-745. 2007, Tom Misteli et al. Molecular Cell Biology. 10: 2
43-254. 2009).
Therefore, if this experimental result is interpreted in a more general manner, it is natural that if a similar experiment is performed on an ATR having a role very similar to ATM in a mammalian cell, AT
Those skilled in the fields of basic medicine, pharmacology, and biochemistry have suggested that R-deficient pigment stem cells will cause differentiation into pigment cells and cause earlier depigmentation compared to wild type. It is clear that we are sure. That is, according to this experiment, ATR or AT
It can be said that the R mutant is also proved to have the function of maintaining and promoting the undifferentiation of the pigment stem cells, similarly to ATM or ATM mutant. Therefore, it is clear that the agent itself containing them promotes the maintenance of undifferentiation of pigment stem cells. Other points are the same as those in the first embodiment, and are not repeated.

<Embodiment 6: Undifferentiated maintenance promoter containing ATR or ATR variant expression vector>
Further, in this embodiment, a promoter sequence that controls ATR or a variant thereof to be expressed in a region containing pigment stem cells, and a downstream of the promoter sequence are linked.
A vector having a nucleotide sequence encoding ATR or an ATR variant,
It is an agent for maintaining the undifferentiation of pigment stem cells. As shown in the examples described later, ATR or ATR mutants were also detected as ATM or ATM by experiments using mice lacking the ATM gene.
It can be said that it has been proved that it has the function of maintaining and promoting the undifferentiation of pigment stem cells as well as the mutant. Therefore, it is clear that an agent itself containing a vector having a nucleotide sequence encoding them also promotes the maintenance of undifferentiation of pigment stem cells. Other points are the same as those in the second embodiment, and will not be repeated.

<Embodiment 7: Self-replication maintenance promoter containing ATR or ATR variant>
This embodiment is an agent for maintaining the self-renewal of pigment stem cells, which comprises an ATR variant having an ATR or pigment cell undifferentiation maintenance promoting effect. As shown in the Examples below, the present inventors have also found mice lacking the ATM gene, which is a major mediator of the genomic damage response,
We are investigating the effects of radiation In general, for genomic damage response, ATM, ATR, H2A
It is known that various proteins such as X (γ-H2AX) and 53BP1 are activated, and these protein groups cooperate to repair damaged DNA (J. Wada Harper et al. Molecul
ar Cell. 28: 739-745. 2007, Tom Misteli et al. Molecular Cell Biology. 10: 243-254
2009). Therefore, by using an ATM-deficient mouse which is a kind of these protein groups, it is possible to examine the possibility that ectopic differentiation of pigment stem cells is performed as part of the genomic damage response. As a result, in pigment stem cells lacking ATM, compared to wild type,
It has been found that it causes differentiation into pigment cells more sensitively and causes earlier depigmentation.
Here, both ATM and ATR are members of the PI-3 kinase family of serine-threonine kinases and are structurally similar, and are located between the sensor and the effector in the genomic damage response, and both are central as transducers. (J. Wada Harper et al. Molecular Cell. 28: 739-745. 2007, Tom Miste
li et al. Molecular Cell Biology. 10: 243-254. 2009). In other words, it is well known that the roles of ATM and ATR partially overlap and work together (J. Wada Harper et al. M
olecular Cell. 28: 739-745. 2007, Tom Misteli et al. Molecular Cell Biology. 10: 2
43-254. 2009).
Therefore, if this experimental result is interpreted in a more general way, it is natural that if a similar experiment is performed on an ATR having a role very similar to ATM in a mammalian cell, the ATR-deficient pigment stem cell It is clear that those skilled in the fields of basic medicine, pharmacy and biochemistry will be more sensitive to differentiation into pigmented cells and cause earlier depigmentation compared to wild type. is there. That is, it can be said that this experiment is equivalent to demonstrating that the ATR or ATR mutant has the function of promoting the maintenance of the ability of the pigment stem cells to self-replicate, similarly to the ATM or ATM mutant. Therefore, it is clear that the agent itself containing them promotes the maintenance of the ability of pigment stem cells to self-replicate. Other points are the same as those in the third embodiment, and are not repeated.

<Embodiment 8: Self-replication maintenance promoter containing ATR or ATR variant expression vector>
In the present embodiment, a promoter sequence that controls expression of ATR or a variant thereof in a region containing pigment stem cells and an ATR linked downstream of the promoter sequence
Alternatively, it is an agent for maintaining the self-renewal of a pigment stem cell, comprising a vector having a nucleotide sequence encoding an ATR variant. As shown in the Examples described later, the function of promoting the maintenance of the ability of pigment stem cells to self-replicate in the same way as ATM or ATM mutants by experiments using mice deficient in ATM genes. Is equal to having been demonstrated to have Therefore, it is clear that the agent itself including the vector having the nucleotide sequence encoding them promotes the maintenance of the ability of pigment stem cells to self-replicate. Other points are the same as those in the fourth embodiment, and are not repeated.

<Embodiments 5 to 8: Description of ATR and ATR variants>
In common with the above embodiments 5 to 8, the amino acid sequence of the human ATR protein is known,
SEQ ID NO: 3 (NCBI Reference Sequence: NP_001175.2 ataxia telangiectasia and
Rad3 related protein [Homo sapiens]).

Sequence number 3: It describes from N terminal to C terminal (2644aa)
1 mgehglelas mipalrelgs atpeeyntvv qkprqilcqf idriltdvnv vavelvkktd
61 sqptsvmlld fiqhimkssp lmfvnvsgsh eakgsciefs nwiitrllri aatpschllh
121 kkicevicsl lflfkskspa ifgvltkell qlfedlvylh rrnvmghave wpvvmsrfls
181 qldehmgylq saplqlmsmq nlefievtll mvltriiaiv ffrrqelllw qigcvlleyg
241 spkikslais fltelfqlgg lpaqpastff ssflellkhl vemdtdqlkl yeeplsklik
301 tlfpfeaeay rniepvylnm lleklcvmfe dgvlmrlksd llkaalchll qyflkfvpag
361 yesalqvrkv yvrnickall dvlgievdae yllgplyaal kmesmeiiee iqcqtqqenl
421 ssnsdgispk rrrlssslnp skrapkqtee ikhvdmnqks ilwsalkqka eslqisleys
481 glknpvieml egiavvlqlt alctvhcshq nmncrtfkdc qhkskkkpsv vitwmsldfy
541 tkvlkscrsl lesvqkldle atidkvvkiy daliymqvns sfedhiledl cgmlslpwiy
601 shsddgclkl ttfaanlltl scrisdsysp qaqsrcvfll tlfprrifle wrtavynwal
661 qssheviras cvsgffillq qqnscnrvpk ilidkvkdds divkkefasi lgqlvctlhg
721 mfyltsslte pfsehghvdl fcrnlkatsq hecsssqlka svckpflfll kkkipspvkl
781 afidnlhhlc khldfredet dvkavlgtll nlmedpdkdv rvafsgnikh ilesldsedg
841 fikelfvlrm keaythaqis rnnelkdtli lttgdigraa kgdlvpfall hllhcllsks
901 asvsgaayte iralvaaksv klqsffsqyk kpicqflves lhssqmtalp ntpcqnadvr
961 kqdvahqrem alntlseian vfdfpdlnrf ltrtlqvllp dlaakaspaa salirtlgkq
1021 lnvnrreili nnfkyifshl vcscskdele ralhylknet eielgsllrq dfqglhnell
1081 lrigehyqqv fnglsilasf assddpyqgp rdiispelma dylqpkllgi laffnmqlls
1141 ssvgiedkkm alnslmslmk lmgpkhvssv rvkmmttlrt glrfkddfpe lccrawdcfv
1201 rcldhaclgs llshvivall plihiqpket aaifhyliie nrdavqdflh eiyflpdhpe
1261 lkkikavlqe yrketsestd lqttlqlsmk aiqhenvdvr ihaltslket lyknqeklik
1321 yatdsetvep iisqlvtvll kgcqdansqa rllcgeclge lgaidpgrld fsttetqgkd
1381 ftfvtgveds sfaygllmel traylayadn sraqdsaaya iqellsiydc remetngpgh
1441 qlwrrfpehv reilephlnt rykssqkstd wsgvkkpiyl sklgsnfaew saswagylit
1501 kvrhdlaski ftccsimmkh dfkvtiyllp hilvyvllgc nqedqqevya eimavlkhdd
1561 qhtintqdia sdlcqlstqt vfsmldhltq warhkfqalk aekcphsksn rnkvdsmvst
1621 vdyedyqsvt rfldlipqdt lavasfrska ytravmhfes fitekkqniq ehlgflqkly
1681 aamhepdgva gvsairkaep slkeqilehe slgllrdata cydraiqlep dqiihyhgvv
1741 ksmlglgqls tvitqvngvh anrsewtdel ntyrveaawk lsqwdlveny laadgksttw
1801 svrlgqllls akkrditafy dslklvraeq ivplsaasfe rgsyqrgyey ivrlhmlcel
1861 ehsikplfqh spgdssqeds lnwvarlemt qnsyrakepi lalrrallsl nkrpdynemv
1921 gecwlqsarv arkaghhqta ynallnages rlaelyvera kwlwskgdvh qalivlqkgv
1981 elcfpenetp pegknmlihg ramllvgrfm eetanfesna imkkykdvta clpewedghf
2041 ylakyydklm pmvtdnkmek qgdliryivl hfgrslqygn qfiyqsmprm ltlwldygtk
2101 ayewekagrs drvqmrndlg kinkviteht nylapyqflt afsqlisric hshdevfvvl
2161 meiiakvfla ypqqamwmmt avskssypmr vnrckeilnk aihmkkslek fvgdatrltd
2221 kllelcnkpv dgssstlsms thfkmlkklv eeatfseili plqsvmiptl psilgthanh
2281 ashepfpghw ayiagfddmv eilaslqkpk kislkgsdgk fyimmckpkd dlrkdcrlme
2341 fnslinkclr kdaesrrrel hirtyavipl ndecgiiewv nntaglrpil tklykekgvy
2401 mtgkelrqcm lpksaalsek lkvfrefllp rhppifhewf lrtfpdptsw yssrsaycrs
2461 tavmsmvgyi lglgdrhgen ilfdsltgec vhvdfnclfn kgetfevpei vpfrlthnmv
2521 ngmgpmgteg lfrracevtm rlmrdqrepl msvlktflhd plvewskpvk ghskaplnet
2581 gevvnekakt hvldieqrlq gviktrnrvt glplsieghv hyliqeatde nllcqmylgw
2641 tpym //

Similarly, the base sequence of the human ATR gene is also known, and the base sequence shown in SEQ ID NO: 4 below (NCBI Reference Sequence: NM_001184.3 Homo sapiens ataxia telangiectasia
and Rad3 related (ATR), mRNA).

SEQ ID NO: 4: described from 5 ′ end to 3 ′ end (8258 bp mRNA)
1 ttccgggagg agttttggcc tccacacggc tccgtcgggc gccgcgctct tccggcagcg
61 gtagctttgg agacgccggg aacccgcgtt ggcgtggttg actagtgcct cgcagcctca
121 gcatggggga acatggcctg gagctggctt ccatgatccc cgccctgcgg gagctgggca
181 gtgccacacc agaggaatat aatacagttg tacagaagcc aagacaaatt ctgtgtcaat
241 tcattgaccg gatacttaca gatgtaaatg ttgttgctgt agaacttgta aagaaaactg
301 actctcagcc aacctccgtg atgttgcttg atttcatcca gcatatcatg aaatcctccc
361 cacttatgtt tgtaaatgtg agtggaagcc atgaggccaa aggcagttgt attgaattca
421 gtaattggat cataacgaga cttctgcgga ttgcagcaac tccctcctgt catttgttac
481 acaagaaaat ctgtgaagtc atctgttcat tattatttct ttttaaaagc aagagtcctg
541 ctatttttgg ggtactcaca aaagaattat tacaactttt tgaagacttg gtttacctcc
601 atagaagaaa tgtgatgggt catgctgtgg aatggccagt ggtcatgagc cgatttttaa
661 gtcaattaga tgaacacatg ggatatttac aatcagctcc tttgcagttg atgagtatgc
721 aaaatttaga atttattgaa gtcactttat taatggttct tactcgtatt attgcaattg
781 tgttttttag aaggcaagaa ctcttacttt ggcagatagg ttgtgttctg ctagagtatg
841 gtagtccaaa aattaaatcc ctagcaatta gctttttaac agaacttttt cagcttggag
901 gactaccagc acaaccagct agcacttttt tcagctcatt tttggaatta ttaaaacacc
961 ttgtagaaat ggatactgac caattgaaac tctatgaaga gccattatca aagctgataa
1021 agacactatt tccctttgaa gcagaagctt atagaaatat tgaacctgtc tatttaaata
1081 tgctgctgga aaaactctgt gtcatgtttg aagacggtgt gctcatgcgg cttaagtctg
1141 atttgctaaa agcagctttg tgccatttac tgcagtattt ccttaaattt gtgccagctg
1201 ggtatgaatc tgctttacaa gtcaggaagg tctatgtgag aaatatttgt aaagctcttt
1261 tggatgtgct tggaattgag gtagatgcag agtacttgtt gggcccactt tatgcagctt
1321 tgaaaatgga aagtatggaa atcattgagg agattcaatg ccaaactcaa caggaaaacc
1381 tcagcagtaa tagtgatgga atatcaccca aaaggcgtcg tctcagctcg tctctaaacc
1441 cttctaaaag agcaccaaaa cagactgagg aaattaaaca tgtggacatg aaccaaaaga
1501 gcatattatg gagtgcactg aaacagaaag ctgaatccct tcagatttcc cttgaataca
1561 gtggcctaaa gaatcctgtt attgagatgt tagaaggaat tgctgttgtc ttacaactga
1621 ctgctctgtg tactgttcat tgttctcatc aaaacatgaa ctgccgtact ttcaaggact
1681 gtcaacataa atccaagaag aaaccttctg tagtgataac ttggatgtca ttggattttt
1741 acacaaaagt gcttaagagc tgtagaagtt tgttagaatc tgttcagaaa ctggacctgg
1801 aggcaaccat tgataaggtg gtgaaaattt atgatgcttt gatttatatg caagtaaaca
1861 gttcatttga agatcatatc ctggaagatt tatgtggtat gctctcactt ccatggattt
1921 attcccattc tgatgatggc tgtttaaagt tgaccacatt tgccgctaat cttctaacat
1981 taagctgtag gatttcagat agctattcac cacaggcaca atcacgatgt gtgtttcttc
2041 tgactctgtt tccaagaaga atattccttg agtggagaac agcagtttac aactgggccc
2101 tgcagagctc ccatgaagta atccgggcta gttgtgttag tggatttttt atcttattgc
2161 agcagcagaa ttcttgtaac agagttccca agattcttat agataaagtc aaagatgatt
2221 ctgacattgt caagaaagaa tttgcttcta tacttggtca acttgtctgt actcttcacg
2281 gcatgtttta tctgacaagt tctttaacag aacctttctc tgaacacgga catgtggacc
2341 tcttctgtag gaacttgaaa gccacttctc aacatgaatg ttcatcttct caactaaaag
2401 cttctgtctg caagccattc cttttcctac tgaaaaaaaa aatacctagt ccagtaaaac
2461 ttgctttcat agataatcta catcatcttt gtaagcatct tgattttaga gaagatgaaa
2521 cagatgtaaa agcagttctt ggaactttat taaatttaat ggaagatcca gacaaagatg
2581 ttagagtggc ttttagtgga aatatcaagc acatattgga atccttggac tctgaagatg
2641 gatttataaa ggagcttttt gtcttaagaa tgaaggaagc atatacacat gcccaaatat
2701 caagaaataa tgagctgaag gataccttga ttcttacaac aggggatatt ggaagggccg
2761 caaaaggaga tttggtacca tttgcactct tacacttatt gcattgtttg ttatccaagt
2821 cagcatctgt ctctggagca gcatacacag aaattagagc tctggttgca gctaaaagtg
2881 ttaaactgca aagttttttc agccagtata agaaacccat ctgtcagttt ttggtagaat
2941 cccttcactc tagtcagatg acagcacttc cgaatactcc atgccagaat gctgacgtgc
3001 gaaaacaaga tgtggctcac cagagagaaa tggctttaaa tacgttgtct gaaattgcca
3061 acgttttcga ctttcctgat cttaatcgtt ttcttactag gacattacaa gttctactac
3121 ctgatcttgc tgccaaagca agccctgcag cttctgctct cattcgaact ttaggaaaac
3181 aattaaatgt caatcgtaga gagattttaa taaacaactt caaatatatt ttttctcatt
3241 tggtctgttc ttgttccaaa gatgaattag aacgtgccct tcattatctg aagaatgaaa
3301 cagaaattga actggggagc ctgttgagac aagatttcca aggattgcat aatgaattat
3361 tgctgcgtat tggagaacac tatcaacagg tttttaatgg tttgtcaata cttgcctcat
3421 ttgcatccag tgatgatcca tatcagggcc cgagagatat catatcacct gaactgatgg
3481 ctgattattt acaacccaaa ttgttgggca ttttggcttt ttttaacatg cagttactga
3541 gctctagtgt tggcattgaa gataagaaaa tggccttgaa cagtttgatg tctttgatga
3601 agttaatggg acccaaacat gtcagttctg tgagggtgaa gatgatgacc acactgagaa
3661 ctggccttcg attcaaggat gattttcctg aattgtgttg cagagcttgg gactgctttg
3721 ttcgctgcct ggatcatgct tgtctgggct cccttctcag tcatgtaata gtagctttgt
3781 tacctcttat acacatccag cctaaagaaa ctgcagctat cttccactac ctcataattg
3841 aaaacaggga tgctgtgcaa gattttcttc atgaaatata ttttttacct gatcatccag
3901 aattaaaaaa gataaaagcc gttctccagg aatacagaaa ggagacctct gagagcactg
3961 atcttcagac aactcttcag ctctctatga aggccattca acatgaaaat gtcgatgttc
4021 gtattcatgc tcttacaagc ttgaaggaaa ccttgtataa aaatcaggaa aaactgataa
4081 agtatgcaac agacagtgaa acagtagaac ctattatctc acagttggtg acagtgcttt
4141 tgaaaggttg ccaagatgca aactctcaag ctcggttgct ctgtggggaa tgtttagggg
4201 aattgggggc gatagatcca ggtcgattag atttctcaac aactgaaact caaggaaaag
4261 attttacatt tgtgactgga gtagaagatt caagctttgc ctatggatta ttgatggagc
4321 taacaagagc ttaccttgcg tatgctgata atagccgagc tcaagattca gctgcctatg
4381 ccattcagga gttgctttct atttatgact gtagagagat ggagaccaac ggcccaggtc
4441 accaattgtg gaggagattt cctgagcatg ttcgggaaat actagaacct catctaaata
4501 ccagatacaa gagttctcag aagtcaaccg attggtctgg agtaaagaag ccaatttact
4561 taagtaaatt gggtagtaac tttgcagaat ggtcagcatc ttgggcaggt tatcttatta
4621 caaaggttcg acatgatctt gccagtaaaa ttttcacctg ctgtagcatt atgatgaagc
4681 atgatttcaa agtgaccatc tatcttcttc cacatattct ggtgtatgtc ttactgggtt
4741 gtaatcaaga agatcagcag gaggtttatg cagaaattat ggcagttcta aagcatgacg
4801 atcagcatac cataaatacc caagacattg catctgatct gtgtcaactc agtacacaga
4861 ctgtgttctc catgcttgac catctcacac agtgggcaag gcacaaattt caggcactga
4921 aagctgagaa atgtccacac agcaaatcaa acagaaataa ggtagactca atggtatcta
4981 ctgtggatta tgaagactat cagagtgtaa cccgttttct agacctcata ccccaggata
5041 ctctggcagt agcttccttt cgctccaaag catacacacg agctgtaatg cactttgaat
5101 catttattac agaaaagaag caaaatattc aggaacatct tggattttta cagaaattgt
5161 atgctgctat gcatgaacct gatggagtgg ccggagtcag tgcaattaga aaggcagaac
5221 catctctaaa agaacagatc cttgaacatg aaagccttgg cttgctgagg gatgccactg
5281 cttgttatga cagggctatt cagctagaac cagaccagat cattcattat catggtgtag
5341 taaagtccat gttaggtctt ggtcagctgt ctactgttat cactcaggtg aatggagtgc
5401 atgctaacag gtccgagtgg acagatgaat taaacacgta cagagtggaa gcagcttgga
5461 aattgtcaca gtgggatttg gtggaaaact atttggcagc agatggaaaa tctacaacat
5521 ggagtgtcag actgggacag ctattattat cagccaaaaa aagagatatc acagcttttt
5581 atgactcact gaaactagtg agagcagaac aaattgtacc tctttcagct gcaagctttg
5641 aaagaggctc ctaccaacga ggatatgaat atattgtgag attgcacatg ttatgtgagt
5701 tggagcatag catcaaacca cttttccagc attctccagg tgacagttct caagaagatt
5761 ctctaaactg ggtagctcga ctagaaatga cccagaattc ctacagagcc aaggagccta
5821 tcctggctct ccggagggct ttactaagcc tcaacaaaag accagattac aatgaaatgg
5881 ttggagaatg ctggctgcag agtgccaggg tagctagaaa ggctggtcac caccagacag
5941 cctacaatgc tctccttaat gcaggggaat cacgactcgc tgaactgtac gtggaaaggg
6001 caaagtggct ctggtccaag ggtgatgttc accaggcact aattgttctt caaaaaggtg
6061 ttgaattatg ttttcctgaa aatgaaaccc cacctgaggg taagaacatg ttaatccatg
6121 gtcgagctat gctactagtg ggccgattta tggaagaaac agctaacttt gaaagcaatg
6181 caattatgaa aaaatataag gatgtgaccg cgtgcctgcc agaatgggag gatgggcatt
6241 tttaccttgc caagtactat gacaaattga tgcccatggt cacagacaac aaaatggaaa
6301 agcaaggtga tctcatccgg tatatagttc ttcattttgg cagatctcta caatatggaa
6361 atcagttcat atatcagtca atgccacgaa tgttaactct atggcttgat tatggtacaa
6421 aggcatatga atgggaaaaa gctggccgct ccgatcgtgt acaaatgagg aatgatttgg
6481 gtaaaataaa caaggttatc acagagcata caaactattt agctccatat caatttttga
6541 ctgctttttc acaattgatc tctcgaattt gtcattctca cgatgaagtt tttgttgtct
6601 tgatggaaat aatagccaaa gtatttctag cctatcctca acaagcaatg tggatgatga
6661 cagctgtgtc aaagtcatct tatcccatgc gtgtgaacag atgcaaggaa atcctcaata
6721 aagctattca tatgaaaaaa tccttagaga agtttgttgg agatgcaact cgcctaacag
6781 ataagcttct agaattgtgc aataaaccgg ttgatggaag tagttccaca ttaagcatga
6841 gcactcattt taaaatgctt aaaaagctgg tagaagaagc aacatttagt gaaatcctca
6901 ttcctctaca atcagtcatg atacctacac ttccatcaat tctgggtacc catgctaacc
6961 atgctagcca tgaaccattt cctggacatt gggcctatat tgcagggttt gatgatatgg
7021 tggaaattct tgcttctctt cagaaaccaa agaagatttc tttaaaaggc tcagatggaa
7081 agttctacat catgatgtgt aagccaaaag atgacctgag aaaggattgt agactaatgg
7141 aattcaattc cttgattaat aagtgcttaa gaaaagatgc agagtctcgt agaagagaac
7201 ttcatattcg aacatatgca gttattccac taaatgatga atgtgggatt attgaatggg
7261 tgaacaacac tgctggtttg agacctattc tgaccaaact atataaagaa aagggagtgt
7321 atatgacagg aaaagaactt cgccagtgta tgctaccaaa gtcagcagct ttatctgaaa
7381 aactcaaagt attccgagaa tttctcctgc ccaggcatcc tcctattttt catgagtggt
7441 ttctgagaac attccctgat cctacatcat ggtacagtag tagatcagct tactgccgtt
7501 ccactgcagt aatgtcaatg gttggttata ttctggggct tggagaccgt catggtgaaa
7561 atattctctt tgattctttg actggtgaat gcgtacatgt agatttcaat tgtcttttca
7621 ataagggaga aacctttgaa gttccagaaa ttgtgccatt tcgcctgact cataatatgg
7681 ttaatggaat gggtcctatg ggaacagagg gtctttttcg aagagcatgt gaagttacaa
7741 tgaggctgat gcgtgatcag cgagagcctt taatgagtgt cttaaagact tttctacatg
7801 atcctcttgt ggaatggagt aaaccagtga aagggcattc caaagcgcca ctgaatgaaa
7861 ctggagaagt tgtcaatgaa aaggccaaga cccatgttct tgacattgag cagcgactac
7921 aaggtgtaat caagactcga aatagagtga caggactgcc gttatctatt gaaggacatg
7981 tgcattacct tatacaggaa gctactgatg aaaacttact atgccagatg tatcttggtt
8041 ggactccata tatgtgaaat gaaattatgt aaaagaatat gttaataatc taaaagtaat
8101 gcatttggta tgaatctgtg gttgtatctg ttcaattcta aagtacaaca taaatttacg
8161 ttctcagcaa ctgttatttc tctctgatca ttaattatat gtaaaataat atacattcag
8221 ttattaagaa ataaactgct ttcttaatac aaaaaaaa //

(1) Specific Examples of Embodiments 5 and 7 (Polypeptide) Here, the ATR or ATR variant described above includes (a) ATR consisting of the amino acid sequence represented by SEQ ID NO: 3, (b) SEQ ID NO: 3. An ATR variant comprising an amino acid sequence having at least 80% homology with the amino acid sequence represented, and having an action to promote undifferentiation or self-renewal of pigment stem cells; (c) a nucleotide represented by SEQ ID NO: 3 ATR encoded by the sequence, or (d) a nucleic acid that hybridizes under stringent conditions to the nucleotide sequence represented by SEQ ID NO: 3 or its complementary strand, and promotes the maintenance of undifferentiation of pigment stem cells or self-replication Any of ATR variants having a maintenance promoting effect is included.

Thus, it is demonstrated that the amino acid sequence having at least 80% homology with the amino acid sequence represented by SEQ ID NO: 3 maintains and promotes the undifferentiation or self-renewal ability of pigment stem cells in the examples described later. In the same way as the ATR consisting of the amino acid sequence represented by SEQ ID NO: 3, it is possible for those skilled in the fields of basic medicine, pharmacology, biochemistry and the like to maintain and promote the undifferentiation or self-renewal ability of pigment stem cells. Is clear. This homology is 8
The content is preferably 0% or more, more preferably 85% or more, still more preferably 90% or more, and most preferably 95% or more. Thus, the higher the homology, the higher the possibility of maintaining and promoting the undifferentiated or self-replicating ability of the pigment stem cells, similar to the ATR consisting of the amino acid sequence represented by SEQ ID NO: 3.

Alternatively, even if the ATR mutant has an amino acid sequence in which one or several amino acid residues are deleted, substituted, or added to the amino acid sequence represented by SEQ ID NO: 3, Similar to the ATR consisting of the amino acid sequence represented by SEQ ID NO: 3, which has been proved to maintain and promote differentiation or self-renewal ability, maintaining and promoting undifferentiation or self-renewal ability of pigment stem cells, It will be apparent to those skilled in the fields of basic medicine, pharmacy, biochemistry and the like. The term “several amino acid residues” as used herein means 2 to 9 amino acid residues, particularly preferably 2 to 5 amino acid residues, and more preferably 2 to 3 amino acid residues. Most preferred. In this way, amino acid residue deletion / substitution /
The smaller the number of additions, the ATR consisting of the amino acid sequence represented by SEQ ID NO: 3.
In the same manner, the possibility of maintaining and promoting the undifferentiated or self-replicating ability of the pigment stem cells is increased.

On the other hand, an AT having an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions to the nucleotide sequence represented by SEQ ID NO: 4 or its complementary strand as described above.
Similar to the ATR encoded by the nucleotide sequence represented by SEQ ID NO: 4, even if it is an R mutant, it has been demonstrated to maintain and promote the undifferentiation or self-renewal ability of pigment stem cells in the examples described below. It is apparent to those skilled in the fields of basic medicine, pharmacology, biochemistry, and the like to maintain and promote the undifferentiated or self-renewal ability of pigment stem cells. Here, stringent conditions can be easily determined by those skilled in the art and are generally calculated empirically depending on the probe length, washing temperature, and salt concentration. The specific conditions described above can be preferably used.

In addition, as long as it is an ATR mutant having an amino acid sequence encoded by a nucleotide sequence having at least 80% homology with the nucleotide sequence represented by SEQ ID NO: 2, the undifferentiated property of the pigment stem cell in Examples described later Alternatively, as in the ATR encoded by the nucleotide sequence represented by SEQ ID NO: 4, which has been demonstrated to maintain and promote the self-renewal ability, it is fundamental to maintain and promote the undifferentiated or self-renewal ability of pigment stem cells. It will be apparent to those skilled in the fields of medicine, pharmacy, biochemistry and the like. The homology is preferably 80% or more, more preferably 85% or more, and still more preferably 90% or more.
Most preferably, it is 95% or more. Thus, the higher the homology, the more SEQ ID NO: 2
In the same manner as the ATR encoded by the nucleotide sequence represented by the formula (2), the possibility of maintaining and promoting the undifferentiated or self-replicating ability of the pigment stem cell is increased.

In addition, even in the case of an ATR variant encoded by a nucleotide sequence in which one or several nucleotide residues are deleted, substituted, or added to the nucleotide sequence represented by SEQ ID NO: 4, examples described later ATR encoded by the nucleotide sequence represented by SEQ ID NO: 4 which has been demonstrated to maintain and promote undifferentiated or self-renewing ability of pigment stem cells
Similarly, it is apparent to those skilled in the fields of basic medicine, pharmacology, biochemistry, etc. that the pigment stem cells are maintained and promoted to be undifferentiated or self-replicating. In addition, although several nucleotide residues here mean 2 or more and 9 or less nucleotide residues, It is especially preferable that they are 2 or more and 5 or less nucleotide residues, and it is 2 or more and 3 or less nucleotide residues. Is most preferred. Thus, the smaller the number of nucleotide residue deletions / substitutions / additions,
Like the ATR encoded by the nucleotide sequence represented by SEQ ID NO: 4, there is a high possibility of promoting the maintenance of undifferentiated or self-replicating ability of the pigment stem cells.

<Embodiment 9: Undifferentiated maintenance promoter containing protein related to DDR>
The present embodiment is an agent for maintaining the undifferentiation of pigment stem cells, which comprises one or more proteins included in a protein group activated by a genome damage inducer. As shown in the Examples described later, the present inventors are examining the influence of irradiation on mice deficient in the ATM gene, which is a major mediator of the genomic damage response. In general, in the genome damage response, ATM, ATR,
It is known that various proteins such as H2AX (γ-H2AX) and 53BP1 are activated and these proteins cooperate to repair damaged DNA (J. Wada Harper et al. M
olecular Cell. 28: 739-745. 2007, Tom Misteli et al. Molecular Cell Biology. 10: 2
43-254. 2009). Therefore, by using an ATM-deficient mouse which is a kind of these protein groups, it is possible to examine the possibility that ectopic differentiation of pigment stem cells is performed as part of the genomic damage response. As a result, it was found that pigment stem cells lacking ATM cause differentiation into pigment cells more sensitively and cause earlier depigmentation compared to wild type. And, if the result of this experiment is generalized and interpreted, it goes without saying that if a similar experiment is performed on one or more proteins included in the protein group activated by the genome damage inducer, the one or more proteins Those skilled in the fields of basic medicine, pharmacology, and biochemistry will be able to induce differentiation into pigment cells and cause earlier depigmentation compared to wild type. It is clear that we are sure. That is, according to this experiment, one or more proteins included in the protein group activated by the genomic damage-inducing agent also have a function of maintaining and promoting chromogenic stem cell undifferentiation, like ATM or ATM variants. It can be said that is equivalent to that demonstrated. Therefore, it is clear that the agent itself containing them promotes the maintenance of undifferentiation of pigment stem cells. For other points,
Since it is the same as that of Embodiment 1, it does not repeat.

Here, the protein activated by the genomic damage response is not particularly limited.
Harper et al. Molecular Cell. 28: 739-745. 2007, Tom Misteli et al. Molecular Ce
ll Biology. 10: 243-254. 2009. Many arbitrary proteins can be mentioned. For example, for proteins such as ATM, ATR, H2AX (γ-H2AX), 53BP1, etc. This is particularly preferred because a number of powerful experimental results have been reported that demonstrate that it is activated by a damage response.

Here, it is known that ATM kinase is involved in activation of a substrate protein by phosphorylating a substrate protein having an SQ / TQ motif. Proteins with SQ / TQ motifs include Chk1 / Chk2 kinase that mediates ATM signals, cell cycle checkpoint and p53 involved in apoptosis, BRC involved in DNA repair
A1 and FANCD2 are known, and it can be said that this system is controlled by phosphorylation of the SQ / TQ motif. Various mediators with SQ / TQ motifs are DNA
AT forms by accumulating at the site of damage and forming a point-like structure called the intranuclear focus.
It is involved in signal transduction by M. H, a kind of histone at the site of DNA damage
When 2AX is phosphorylated, MDC1 accumulates at this site, followed by NBS1, BRC
A1, 53BP1, FANCD2, Chk2, etc. are recruited. MDC1, NBS1
, BRCA1 and 53BP1 are adapter proteins having a BRCT domain, and these are considered to function as scaffolds for efficiently collecting various signal molecules at a damaged site and performing DNA repair and phosphorylation by ATM efficiently. Therefore, these proteins are also preferably used in the present embodiment as proteins that are activated by a genomic damage response.

<Embodiment 10: Undifferentiated maintenance promoter containing an expression vector for a protein related to DDR>
In addition, the present embodiment is a promoter sequence that is controlled to be expressed in a region containing a pigment stem cell, and one or more proteins included in a group of proteins that are linked downstream of the promoter sequence and activated by a genome damage inducer. And a non-differentiated maintenance promoter for pigment stem cells, comprising a vector having a nucleotide sequence encoding the protein of As shown in the examples described later, one or more proteins included in the protein group activated by the genome damage-inducing agent by an experiment using a mouse deficient in the ATM gene, as well as ATM or ATM variants,
Equivalent to having the function of promoting the maintenance of the ability of pigment stem cells to self-renew. Therefore, it is clear that an agent itself containing a vector having a nucleotide sequence encoding them also promotes the maintenance of undifferentiation of pigment stem cells. For other points,
Since it is the same as that of Embodiment 2, it does not repeat.

<Embodiment 11: Undifferentiated maintenance promoter containing a control factor related to DDR>
The present embodiment is a pigment stem cell undifferentiation maintenance promoter containing a regulatory factor that controls the activity of one or more proteins contained in a protein group activated by a genome damage inducer. As shown in the Examples described later, the present inventors are examining the influence of irradiation on mice deficient in the ATM gene, which is a major mediator of the genomic damage response. In general, it is known that various proteins such as ATM, ATR, H2AX (γ-H2AX), and 53BP1 are activated in genome damage response, and these proteins are cooperated to repair damaged DNA. (J. Wada Harper et al. Molecular Cell. 28: 739-745. 2007, Tom Misteli et al. Mole
cular Cell Biology. 10: 243-254. 2009). Therefore, A which is a kind of these protein groups
By using TM-deficient mice, it is possible to examine the possibility that ectopic differentiation of pigment stem cells is performed as part of the genomic damage response. As a result, it was found that pigment stem cells lacking ATM cause differentiation into pigment cells more sensitively and cause earlier depigmentation compared to wild type. And, if the result of this experiment is generalized and interpreted, it goes without saying that if a similar experiment is performed on one or more proteins included in the protein group activated by the genome damage inducer, the one or more proteins Those skilled in the fields of basic medicine, pharmacology, and biochemistry will be able to induce differentiation into pigment cells and cause earlier depigmentation compared to wild type. It is clear that we are sure. As a result, as a matter of course, it is clear that the agent itself containing a regulatory factor that controls their activity also maintains and promotes the undifferentiation of pigment stem cells. That is, according to this experiment, one or more proteins included in the protein group activated by the genomic damage-inducing agent also have a function of maintaining and promoting chromogenic stem cell undifferentiation, like ATM or ATM variants. It can be said that is equivalent to that demonstrated. Therefore, it is clear that agents containing regulatory factors that control their activity also maintain and promote the undifferentiation of pigment stem cells.

Here, the protein activated by the genomic damage response is not particularly limited.
Harper et al. Molecular Cell. 28: 739-745. 2007, Tom Misteli et al. Molecular Ce
ll Biology. 10: 243-254. 2009 can include a regulatory factor that controls the activity of a protein activated by a genome damage-inducing agent in a positive direction or a negative direction.
Here, it is known that ATM kinase is involved in activation of a substrate protein by phosphorylating a substrate protein having an SQ / TQ motif. Proteins with SQ / TQ motifs include Chk1 / Chk2 kinase that mediates ATM signals, cell cycle checkpoint and p53 involved in apoptosis, BRC involved in DNA repair
A1 and FANCD2 are known, and it can be said that this system is controlled by phosphorylation of the SQ / TQ motif. Various mediators with SQ / TQ motifs are DNA
AT forms by accumulating at the site of damage and forming a point-like structure called the intranuclear focus.
It is involved in signal transduction by M. H, a kind of histone at the site of DNA damage
When 2AX is phosphorylated, MDC1 accumulates at this site, followed by NBS1, BRC
A1, 53BP1, FANCD2, Chk2, etc. are recruited. MDC1, NBS1
, BRCA1 and 53BP1 are adapter proteins having a BRCT domain, and these are considered to function as scaffolds for efficiently collecting various signal molecules at a damaged site and performing DNA repair and phosphorylation by ATM efficiently. Therefore, these proteins are also preferably used in this embodiment as control factors that control the activity of proteins activated by the genomic damage response in a positive direction or a negative direction.
Other points are the same as those in the ninth or tenth embodiment and will not be repeated.

<Embodiment 12: Self-renewal maintenance promoter containing protein related to DDR>
This embodiment is a pigment stem cell self-renewal maintenance promoter containing one or more proteins included in a protein group activated by a genome damage-inducing agent. As shown in the Examples described later, the present inventors are examining the influence of irradiation on mice deficient in the ATM gene, which is a major mediator of the genomic damage response. In general, in the genome damage response, ATM, ATR,
It is known that various proteins such as H2AX (γ-H2AX) and 53BP1 are activated and these proteins cooperate to repair damaged DNA (J. Wada Harper et al. M
olecular Cell. 28: 739-745. 2007, Tom Misteli et al. Molecular Cell Biology. 10: 2
43-254. 2009). Therefore, by using an ATM-deficient mouse which is a kind of these protein groups, it is possible to examine the possibility that ectopic differentiation of pigment stem cells is performed as part of the genomic damage response. As a result, it was found that pigment stem cells lacking ATM cause differentiation into pigment cells more sensitively and cause earlier depigmentation compared to wild type. And, if the result of this experiment is generalized and interpreted, it goes without saying that if a similar experiment is performed on one or more proteins included in the protein group activated by the genome damage inducer, the one or more proteins Those skilled in the fields of basic medicine, pharmacology, and biochemistry will be able to induce differentiation into pigment cells and cause earlier depigmentation compared to wild type. It is clear that we are sure. That is, as a result of this experiment, one or more proteins included in the protein group activated by the genome damage-inducing agent also have the function of promoting the maintenance of the ability of pigment stem cells to self-replicate, as with ATM or ATM variants. This is equivalent to being proven. Therefore, it is clear that the agent itself containing them promotes the maintenance of the ability of pigment stem cells to self-replicate. Other points are the same as those in the third embodiment, and are not repeated.

<Embodiment 12: Self-replication maintenance promoter containing an expression vector for a protein related to DDR>
In the present embodiment, a promoter sequence that is controlled to be expressed in a region including a pigment stem cell, and one or more proteins included in a protein group that is linked downstream of the promoter sequence and activated by a genome damage inducer A self-renewal promoter for pigment stem cells, comprising a vector having a nucleotide sequence encoding As shown in the examples below, A
Through experiments using mice deficient in the TM gene, one or more proteins included in the protein group activated by the genomic damage inducer also maintain the ability of pigment stem cells to self-replicate, as with ATM or ATM variants. Equivalent to having a function of promoting. Therefore, it is clear that the agent itself including the vector having the nucleotide sequence encoding them promotes the maintenance of the ability of pigment stem cells to self-replicate. For other points,
Since it is the same as that of Embodiment 4, it does not repeat.

<Embodiment 13: Self-renewal maintenance promoter containing a control factor related to DDR>
This embodiment is a self-replication promoting agent for pigment stem cells, which contains a regulatory factor that controls the activity of one or more proteins contained in a protein group activated by a genome damage inducer. As shown in the Examples described later, the present inventors are examining the influence of irradiation on mice deficient in the ATM gene, which is a major mediator of the genomic damage response. In general, it is known that various proteins such as ATM, ATR, H2AX (γ-H2AX), and 53BP1 are activated in genome damage response, and these proteins are cooperated to repair damaged DNA. (J. Wada Harper et al. Molecular Cell. 28: 739-745. 2007, Tom Misteli et al. Mole
cular Cell Biology. 10: 243-254. 2009). Therefore, A which is a kind of these protein groups
By using TM-deficient mice, it is possible to examine the possibility that ectopic differentiation of pigment stem cells is performed as part of the genomic damage response. As a result, it was found that pigment stem cells lacking ATM cause differentiation into pigment cells more sensitively and cause earlier depigmentation compared to wild type. And, if the result of this experiment is generalized and interpreted, it goes without saying that if a similar experiment is performed on one or more proteins included in the protein group activated by the genome damage inducer, the one or more proteins Those skilled in the fields of basic medicine, pharmacology, and biochemistry will be able to induce differentiation into pigment cells and cause earlier depigmentation compared to wild type. It is clear that we are sure. As a result, as a matter of course, it is clear that an agent itself containing a regulatory factor that controls their activity also maintains and promotes the self-renewal ability of pigment stem cells. That is, as a result of this experiment, one or more proteins included in the protein group activated by the genomic damage-inducing agent also have the function of promoting the maintenance of the ability of pigment stem cells to self-replicate, as with ATM or ATM variants. Equivalent to being proven. Therefore, it is clear that agents containing regulatory factors that control their activity themselves promote the maintenance of the ability of pigmented stem cells to self-replicate. Other points are the same as in the eleventh and twelfth embodiments, and will not be repeated.

<Embodiment 14: Depigmentation therapeutic agent containing an undifferentiated maintenance promoter>
This embodiment is a depigmenting therapeutic agent for mammals containing the above-described agent for maintaining the differentiation of pigment stem cells. As shown in the Examples below, the above-described pigment stem cell undifferentiation maintenance promoter has a function of maintaining and promoting pigment stem cell undifferentiation, and therefore, a depigmentation therapeutic agent for mammals containing it itself It also promotes maintaining the undifferentiation of pigment stem cells. Thereby, the depigmentation therapeutic agent can suppress depigmentation due to depletion of pigment stem cells.

In the present embodiment, the “depigmentation therapeutic agent” means a medicine having an action of suppressing so-called gray hair (or depigmentation of mammalian hair). Here, “whitening” means a phenomenon in which white hair gradually increases from what originally had black hair. Also,
“Hair” refers to an appendage of the skin consisting of a hair shaft and hair root mainly composed of keratin, and includes, for example, head hair and body hair. And "hair depigmentation" means that the hair that originally has melanin pigment is reduced by the supply of melanin pigment from the pigment cells,
This means that the color of the hair approaches white. This includes, for example, the change of hair to white hair and whitening of body hair.

Here, there is a possibility that the therapeutic agent of the present embodiment has the effect by oral administration or transdermal administration, for example, powder, pill, tablet, capsule, cream, It may be a liquid preparation or an injection. In addition, the content of the active ingredient (the above-mentioned various proteins, vectors, regulatory factors, etc.) of the pigment stem cell undifferentiation maintenance promoter contained in this therapeutic agent is:
Depending on the type of active ingredient and the disease of the patient, the dosage may be adjusted as appropriate at the discretion of a medical professional such as a doctor. Furthermore, the frequency of administration of the therapeutic agent, the administration interval, and the like are similarly appropriately adjusted at the discretion of a medical professional such as a doctor depending on the type of active ingredient and the disease of the patient. In addition,
The description of these therapeutic agents is common to other embodiments.

<Embodiment 15: Depigmentation therapeutic agent containing self-replication inhibition inhibitor>
The present embodiment is a therapeutic agent for depigmentation of a mammal, including the above-described inhibitor of inhibition of pigment stem cell self-renewal. As shown in the Examples below, the above-described pigment stem cell undifferentiation maintenance promoter has a function of promoting the maintenance of the ability of pigment stem cells to self-replicate. As such, it promotes the maintenance of the ability of pigment stem cells to self-renew. This
A depigmentation therapeutic agent can suppress depigmentation due to depletion of pigment stem cells. Other points are the same as in the fourteenth embodiment, and will not be repeated.

<Embodiment 15: Tissue stem cell undifferentiated maintenance promoter containing a protein related to DDR>
This embodiment is an agent for maintaining undifferentiated tissue stem cells, which includes one or more proteins included in a protein group activated by a genome damage inducer. As shown in the Examples described later, the present inventors are examining the influence of irradiation on mice deficient in the ATM gene, which is a major mediator of the genomic damage response. In general, in the genome damage response, ATM, ATR,
It is known that various proteins such as H2AX (γ-H2AX) and 53BP1 are activated and these proteins cooperate to repair damaged DNA (J. Wada Harper et al. M
olecular Cell. 28: 739-745. 2007, Tom Misteli et al. Molecular Cell Biology. 10: 2
43-254. 2009). Therefore, by using an ATM-deficient mouse which is a kind of these protein groups, it is possible to examine the possibility that ectopic differentiation of pigment stem cells is performed as part of the genomic damage response. As a result, it was found that pigment stem cells lacking ATM cause differentiation into pigment cells more sensitively and cause earlier depigmentation compared to wild type. If this experimental result is interpreted in a more general manner, it has been demonstrated that one or more proteins included in the protein group activated by the genomic damage inducer have a function of maintaining and promoting undifferentiation of tissue stem cells. It can be said that Therefore, it is clear that the agent itself containing them promotes the maintenance of undifferentiation of tissue stem cells.

Thus, the present inventors have found that the above-mentioned ectopic differentiation of tissue stem cells is caused when the genome of tissue stem cells is damaged by the genome damage inducer. That is,
Promoting the maintenance of undifferentiated tissue stem cells refers to suppressing the differentiation of tissue stem cells into mature cells caused by such a genome damage-inducing source. In addition, description of the promotion of undifferentiated maintenance of these tissue stem cells is common to other embodiments.

Here, the tissue stem cells other than the pigment stem cells are not particularly limited, and can include a large number of arbitrary tissue stem cells.
This is particularly preferable because powerful experimental results have been reported that demonstrate that proteins related to DDR such as TM and H2AX (γ-H2AX) are activated. Examples of tissue stem cells other than these include blood stem cells, mesenchymal stem cells, neural stem cells, corneal epithelial stem cells, and epidermal stem cells. The description of these tissue stem cells is common to other embodiments. The other points are the same as in the first embodiment, and are not repeated.

<Embodiment 16: An agent for maintaining undifferentiation of tissue stem cells containing an expression vector for a protein related to DDR>
In the present embodiment, a promoter sequence that is controlled to be expressed in a region containing a tissue stem cell, and one or more proteins included in a protein group that is linked downstream of the promoter sequence and activated by a genome damage inducer And an undifferentiated maintenance promoter for tissue stem cells, comprising a vector having a nucleotide sequence encoding As shown in the examples below, A
As a result of experiments using mice deficient in the TM gene, one or more proteins included in the protein group activated by the genomic damage-inducing agent are also undifferentiated of tissue stem cells of tissue stem cells, like ATM or ATM variants. Equivalent to having a function of maintaining and promoting Therefore, it is clear that an agent itself containing a vector having a nucleotide sequence encoding them also promotes maintenance of undifferentiation of tissue stem cells of pigment stem cells. Other points are the same as those in the second embodiment, and will not be repeated.

<Embodiment 17: Undifferentiated maintenance promoter for tissue stem cells containing a regulatory factor related to DDR>
The present embodiment is an agent for maintaining undifferentiated tissue stem cells, which contains a regulatory factor that controls the activity of one or more proteins contained in a protein group activated by a genome damage inducer. As shown in the Examples described later, the present inventors are examining the influence of irradiation on mice deficient in the ATM gene, which is a major mediator of the genomic damage response. In general, it is known that various proteins such as ATM, ATR, H2AX (γ-H2AX), and 53BP1 are activated in genome damage response, and these proteins are cooperated to repair damaged DNA. (J. Wada Harper et al. Molecular Cell. 28: 739-745. 2007, Tom Misteli et al. Mole
cular Cell Biology. 10: 243-254. 2009). Therefore, A which is a kind of these protein groups
By using TM-deficient mice, it is possible to examine the possibility that ectopic differentiation of pigment stem cells is performed as part of the genomic damage response. As a result, it was found that pigment stem cells lacking ATM cause differentiation into pigment cells more sensitively and cause earlier depigmentation compared to wild type. And, if the result of this experiment is generalized and interpreted, it goes without saying that if a similar experiment is performed on one or more proteins included in the protein group activated by the genome damage inducer, the one or more proteins Those of ordinary skill in the fields of basic medicine, pharmacology, and biochemistry will be able to induce differentiation into tissue stem cells more sensitively and earlier maturation differentiation compared to wild type. It is clear to be convinced. As a result, as a matter of course, it is clear that an agent itself containing a regulatory factor that controls their activity also maintains and promotes the undifferentiation of tissue stem cells. That is, according to this experiment, one or more proteins included in the protein group activated by the genome damage-inducing agent also have a function of promoting the maintenance of undifferentiated tissue stem cells, like ATM or ATM variants. Equivalent to being proven. Therefore, it is clear that agents containing regulatory factors that control their activity also promote the maintenance of undifferentiated tissue stem cells. Other points are the same as those in the eleventh embodiment and will not be repeated.

<Embodiment 18: An anti-aging therapeutic agent comprising an agent for maintaining undifferentiated tissue stem cells>
This embodiment is a therapeutic agent for suppressing aging of mammals, which includes the above-described agent for promoting maintenance of undifferentiation of tissue stem cells. As shown in the Examples below, the above-mentioned tissue stem cell undifferentiation maintenance promoter has a function of maintaining and promoting the undifferentiation of tissue stem cells, so that the therapeutic agent for suppressing aging of mammals containing it itself is also included. Maintains and promotes the undifferentiation of tissue stem cells. Thereby, the anti-aging therapeutic agent can suppress the aging phenomenon due to depletion of tissue stem cells.

In the present embodiment, the “aging treatment drug” means a medicine having an action of suppressing so-called aging. Here, “aging” means biologically a change that occurs in an individual organism with the passage of time, and in particular, a functional deterioration that occurs until the organism dies and a phenomenon that occurs in the process. To do. Further, the level of “aging suppression” may suppress any level of aging among the cell level, tissue level, and individual level. For example, aging at the cellular level (cell aging) is known from research using cultured cells. In this aging at the cellular level, when cells removed from living tissue are cultured in vitro, the number of cell divisions is limited, and one reason is that aging progressed because the telomere structure at the end of the chromosome was shortened. . Therefore, for example, it may be effective to evaluate the suppression of aging by using as an index that the telomere structure at the end of the chromosome can be suppressed. However, the index of aging is not limited to this index, and other known markers such as SMP30 that accompany aging, and depending on the case, the amount and quality of tissue stem cells are preferably used as the index. be able to. Even when aging by any index is suppressed, it can be evaluated as having the “aging suppression” action in the present embodiment.

<Embodiment 19: Cosmetic composition comprising a protein related to DDR or a regulator thereof>
This embodiment is a cosmetic composition containing one or more proteins included in a protein group activated by a genome damage inducing agent or a regulator thereof. In this cosmetic composition, as shown in Examples below, one or more proteins included in a protein group activated by a genome damage inducer or a regulator thereof maintain and promote undifferentiation of tissue stem cells. Thus, the depletion due to the differentiation of tissue stem cells is suppressed, and the effect of maintaining youthful hair or bare skin is exhibited.

In the present embodiment, the “cosmetic composition” is a composition that is applied to the face, skin, or hair for cosmetic purposes, and is particularly preferably “ Means "goods". In this embodiment, the cosmetic composition can include at least one other normal cosmetic ingredient in addition to one or more proteins included in the protein group activated by the above-described genome damage inducing agent, Antioxidants, perfumes, preservatives, neutralizers, surfactants, sunscreens, whitening agents, vitamins, wetting agents, self-tanning compounds, anti-wrinkle active agents, emollients, hydrophilic or lipophilic activities Agents, free radical scavengers, deodorants, sequestering agents and film formers and mixtures thereof.

Further, in the present embodiment, the form of the “cosmetic composition” is not particularly limited, and examples thereof include hair styling, hair nourishing, scalp, hair coloring, hair rinsing, hair rinse, lotion,
Cosmetic liquid, cream, milky lotion, sunburn (for), sunscreen (for), cleaning, shaving (for), dead hair sledge (for), facial rinse, pack, cosmetic oil, body rinse, massage (material) ), Foundation, makeup base, funny, lipstick, eye makeup, cheek cosmetics, body makeup, perfume, eau de cologne, bath cosmetics, nail cosmetics, body powders and the like.

For example, in this embodiment, when manufactured and sold in Japan as a cosmetic composition,
False and exaggerated indications recognized under the fair competition rules regarding cosmetics such as “scalp and keep hair healthy”, “grow and apply hair”, “smooth skin” and “keep skin healthy” And cosmetic compositions displayed in a form that does not become. In addition, from the experimental data in Examples described later, one or more proteins included in the protein group activated by the genome damage inducer contained in the cosmetic composition of the present embodiment or a regulator thereof is an undifferentiated tissue stem cell. By maintaining and promoting sex, by suppressing the depletion due to the differentiation of tissue stem cells, "Keep scalp and hair healthy", "Grow and apply hair", "Skin preparation",
It will be apparent to those skilled in the art that it has effects such as “keep the skin healthy”.

<Embodiment 20: Functional food containing a protein related to DDR or a regulator thereof>
This embodiment is a functional food containing one or more proteins included in a protein group activated by a genome damage inducing agent or a regulator thereof. As shown in the examples described later, this functional food is composed of one or more proteins included in the protein group activated by the genome damage inducer or a regulator thereof, as shown in the examples described later. By maintaining and promoting the undifferentiated nature of the skin, it has the effect of maintaining youthful hair, bare skin, or the body environment.

In addition, the form of the functional food of this embodiment is not particularly limited. For example, confectionery shaped like sticks or biscuits, liquid foods, sweeteners, etc. added as a drink In addition, there may be mentioned tablets, capsules, powders and the like.

Further, in the present embodiment, the “functional food” may be any food having some physiologically superior function, and of course, “specific health food (conditions) approved by the Japanese Ministry of Health, Labor and Welfare” (Including food for specified health use) ”and“ functional food products ”such as“ food with functional nutrition ”, but are not limited to these functional food products. In other words, in this embodiment, “functional food” means “health food” that does not require permission from the Japanese Ministry of Health, Labor and Welfare (in addition to the prohibition of false exaggeration labeling of the Health Promotion Act, It is a concept that includes confusing names for functional foods, functions of nutritional components, and indications that specific health objectives can be expected), and those that do not conflict with the Pharmaceutical Affairs Law, the Premium Labeling Law, etc.).
For example, in this embodiment, when producing and selling health foods in Japan, “I want to be a youthful hairstyle”, “I want to be a youthful hairstyle forever”, “I want to be a black-haired Japanese beauty”, etc. Foods that are displayed in a form that does not become a false exaggeration display. In addition,
From the experimental data in the examples described later, one or more proteins included in the protein group activated by the genomic damage inducer contained in the functional food of the present embodiment or its control factor is responsible for the undifferentiation of tissue stem cells. By promoting maintenance, by suppressing the depletion due to differentiation of tissue stem cells, "I want to have a youthful hairstyle forever"
It is considered to be obvious to those skilled in the art that the food is suitable for consumers who have a need such as “I want to be a black-haired Japanese beauty”.

In the present embodiment, “functional food” is a concept including “medical food”, “care food”, and the like, which will be described later. Here, in the present embodiment, the “medical diet” is not particularly limited, but is intended to prevent the progression of the disease or aim for improvement by adjusting the amount of nutrition according to the disease the patient is suffering from. It may be a recuperation meal for a patient who is worried about the deterioration of his / her appearance because, for example, whitening is progressing due to side effects of an anticancer agent or the like. Specific examples of this medical diet include a general medical diet to which one or more proteins included in the protein group activated by the genomic damage-inducing agent or a regulator thereof are added, or a staple diet or a side diet. Examples include those to which proteins have been added.

In the present embodiment, the “care food” is not particularly limited, but is a meal that has been processed so as to be easy to eat so that it can cope with a person with reduced eating and swallowing functions, and ingesting food in a normal form Ingestion and swallowing for those who can not absorb enough nutrition from the ingested food, but elderly people who care about opposite eyes in elderly care facilities and do hair dyeing every day It may be a meal that is easily processed and aims to promote nutrient absorption. Specific examples of this nursing food include those in which one or more proteins included in the protein group activated by the genome damage-inducing factor or its regulatory factor are added to a general nursing food, Proteins added with increased water content compared to normal ones, processed softly, and those with increased viscosity due to the addition of these proteins or their control factors. Can be mentioned.

As mentioned above, although embodiment of this invention was described with reference to drawings, these are the illustrations of this invention, Various structures other than the above are also employable.

For example, the administration target of the agent and the therapeutic agent of the present embodiment is not particularly limited as long as it is a mammal, regardless of gender and age, but is especially prevented when hair depigmentation or progression of aging is concerned. Or therapeutically. In addition, when there is a tendency for hair depigmentation to progress in relatives, the gene of one or more proteins or mutants contained in the protein group that is genetically activated by the genomic damage inducer is deleted. Or the expression of the gene may be poor, and is particularly preferable as a subject of administration.

Furthermore, for these administration subjects, as a result of examining the gene using various well-known techniques in advance, one or more proteins included in the protein group that is actually activated by the genomic damage inducer or It is more preferable to target male humans who are deficient in the mutant gene or whose gene expression has been confirmed to be poor.

EXAMPLES Hereinafter, although an Example demonstrates this invention further, this invention is not limited to these.

(Example 1) Analysis of fate of pigment stem cells by genomic damage inducer (1) Experimental animals Dct-lacZ transgenic mice used in this study were provided by Dr. Ian Jackson, MRC, and XPD mutant mice EMC, provided by Dr. Jan Hoeijmakers.

(2) Ultraviolet irradiation method The mouse is irradiated with ultraviolet rays at 50 kV using Hitachi MBR-1520 (Hitachi Medical).
Using a 2.0 mm aluminum filter with a strength of 20 mA, p, 0.4 Gy / m
Irradiated with a dose of in.

(3) Immunohistochemistry 4% paraformaldehyde fixed on the dorsal skin of the mouse or as it is, OCTcom
After frozen embedding with pound (Sakura Finetechnical), it was cut into 8 μm sections and blocked with 3% skim milk at room temperature for 30 minutes. Then, dilute the primary antibody appropriately to 4 ° C
The mixture was allowed to react overnight, washed with PBS, and then reacted with the secondary antibody.
Primary antibodies used for immunostaining were anti-phosphorylated histone H2AX antibody (Cell Signaling),
Anti-53BP1 antibody (Lifespan BioSciences) and anti-β-galactosidase antibody (Cappel). As the secondary antibody, AlexaFlor 488, 568, 594 (Molecular Probes) was used.

(Results and Discussion)
The present inventors focused on the possibility that the appearance of ectopic pigment cells in the bulge region may be involved in the genomic damage response reaction, and investigated the effect of irradiation on living mice.

First, as shown in FIG. 1, the changes in body color and histology were examined in mice after irradiation. As a result, white hair was induced by irradiating the mouse with 5 Gy of radiation (FIGS. 1a and b). In addition, when histological changes in the bulge region were examined using mice in the growth phase V after irradiation, pigment granules were ectopically formed in a dendritic form instead of undifferentiated pigment stem cells. Pigment cells had appeared (Fig. 1c ~
j).

Further, in the next hair cycle after irradiation, pigment stem cells were not confirmed in the bulge region, and the hair was depigmented (FIGS. 1 k to n). Furthermore, detailed analysis of the hair cycle revealed that pigment cells expressed ectopically in the bulge region were phagocytosed by the keratinocytes surrounding the bulge region and disappeared from the bulge region. It became clear that

From these facts, it is considered that radiation irradiation inhibits pigment stem cell self-replication by inducing differentiation of pigment stem cells in the bulge region, resulting in depletion of pigment stem cells and white hair in the next hair cycle. (FIGS. 1k-o).

In addition, when various chemical substances such as brusphan, mitomycin C, and hydrogen peroxide are administered to mice, differentiation from pigment stem cells to pigment cells is induced in the bulge region as in the case of irradiation, and the next hair cycle is induced. Gray hair was induced in Fig. 1p-w.

In addition, pigment cells induced by the above-mentioned radiation and chemical substances are used in aging mice (22 years old).
It closely resembles the cells confirmed in the bulge region (non-patent document 2, FIG. 1x). Furthermore, in XPD ^{R722W / R · 722W} mice, which are known to express premature traits, pigment cells were confirmed in the bulge region at the mid-growth stage, but not in wild-type mice. It was suggested that even sexual genomic damage causes depigmentation as well (FIGS. 1y, z).

From the above results, pigment stem cells lose their undifferentiated and self-replicating ability due to various types of genomic damage inducers other than irradiation, resulting in depletion of pigment stem cells and depigmentation in the next hair cycle. It has been shown that

Next, in order to investigate whether the genomic damage response was activated in the pigment stem cells after irradiation, the focus formation in the nucleus of phosphorylated H2AX (γ-H2AX), 53BP1 was examined by immunohistochemistry. Note that γ-H2AX and 53BP1 are a type of protein group that is activated by a genome damage inducer.

After irradiation with 5 Gy, γ-H2AX appears not only in the pigment stem cells around the bulge region but also in cells other than the pigment stem cells, and the entire bulge region including the pigment stem cells responds to genomic damage caused by irradiation. (FIGS. 2a-j). Furthermore, the expression of γ-H2AX was maintained for up to 6 hours after irradiation while gradually reducing the number and intensity (FIGS. 2f, h, k, l). In particular, ectopic pigment cells expressed in the bulge region contain γ-H2AX.
Was expressed until the growth stage IV (FIG. 2k, l).

From the above results, it has been clarified that the appearance of ectopic pigment cells in the bulge region is performed as part of the genomic damage response reaction. In addition, it was found that pigment stem cells are destined to differentiate in the bulge region when they suffer from a level of genomic damage that cannot be repaired by irradiation or excessive repair of genomic damage.

That is, since it has been clarified that the appearance of ectopic pigment cells in the bulge region is caused by a protein group activated by a genome damage inducer, in the present embodiment, one or more types included in this protein group are included. It has been clarified that by expressing a protein or controlling the activity of the protein, the differentiation of pigment stem cells is suppressed, the self-replication ability of the pigment stem cells is maintained, and depigmentation of hair can be suppressed.

In addition, it is considered that tissue stem cell differentiation is suppressed and the ability of tissue stem cells to self-replicate is maintained and aging can be suppressed by a mechanism similar to that of pigment stem cells.

(Example 2) Analysis of ATM-deficient mice (1) Experimental animals The ATM-deficient mice used in this study were Peter of St Jude Children's Research Hospital.
J, provided by Dr. McKinnon.

(2) Immunohistochemistry The back skin of mice was immunostained in the same manner as in Example 1. Primary antibodies used for immunostaining were anti-phosphorylated histone H2AX antibody (Cell Signaling), anti-β-galactosidase antibody (
Cappel).

(Results and Discussion)
In order to examine the possibility that the appearance of ectopic pigment cells in the bulge region is involved in the genomic damage response reaction, the present inventors have studied ATM-deficient mice and control mice (
(ATM + / +, ATM +/−) were irradiated and observed for their progress.

First, ATM-deficient mice and control mice were irradiated with 3 Gy radiation, and changes in body color were examined. As a result, in the ATM deficient mice, gray hair was dramatically induced from the first hair cycle after irradiation, whereas in the control mice, white hair was hardly induced even in the second hair cycle after irradiation (FIG. 3a). ).

Next, ATM-deficient mice and control mice were irradiated with 3 Gy radiation, and the expression of γ-H2AX was examined by immunohistochemistry. As described above, γ-H2AX is one type of protein group that is activated in the genome damage response. Therefore, by examining the expression of γ-H2AX in the bulge region, it can be determined whether a genomic damage response is caused in the pigment stem cell.

As a result, at 6 hours after irradiation, γ-H2AX was expressed in all ATM-deficient mice and control mice (FIGS. 3b to d), but at 12 hours after irradiation,
It was found that γ-H2AX was expressed only in ATM-deficient mice (FIGS. 3e-g). That is, it was found that the ATM-deficient mouse caused a long-term genomic damage response compared to the control mouse.

Further, ATM deficient mice and control mice were irradiated with 3 Gy radiation, and the expression of pigment cells in the bulge region 5 days after irradiation was examined by immunohistochemistry. As a result, in ATM-deficient mice, dendritic pigmented pigment cells were observed in the bulge region (FIG. 3j), but pigment cells were not confirmed in control mice (FIG. 3h, i). In addition, it was found that, after irradiation, ectopic differentiation of pigment stem cells was significantly increased in ATM-deficient mice, whereas ectopic differentiation of pigment stem cells was only slightly increased in control mice (FIG. 3k). .

Furthermore, it was revealed that pigment cells induced in ATM-deficient mice are not significantly different from pigment cells that appear when wild-type mice are irradiated with 5 Gy radiation. This
It has been suggested that ATM gene deficiency promotes ectopic differentiation of pigment stem cells in the bulge region, resulting in premature depigmentation.

In particular, even in wild-type mice irradiated with 3 Gy radiation, a dose at which no detectable change can be found in hair color or pigment stem cells, the genomic damage response was remarkably induced. This suggests that it prevents the differentiation and depigmentation of pigment stem cells in the region.

Furthermore, in ATM-deficient mice, a genomic damage response in the bulge region after 3 Gy irradiation was clearly observed over a long period of time. This suggests that in ATM-deficient mice, the efficiency of the genome damage response in the pigment stem cells is reduced, and the resulting damage to the unrepairable genome causes differentiation in the pigment stem cells.

That is, it has been clarified that the appearance of ectopic pigment cells in the bulge region is caused by a protein group activated by a genome damage inducer including ATM. In this embodiment, it is included in this protein group. It became clear that by expressing one or more proteins or controlling the activity of the proteins, the differentiation of the pigment stem cells was suppressed, the self-replication ability of the pigment stem cells was maintained, and the depigmentation of the hair could be suppressed. .

In addition, it is considered that tissue stem cell differentiation is suppressed and the ability of tissue stem cells to self-replicate is maintained and aging can be suppressed by a mechanism similar to that of pigment stem cells.

In the above, this invention was demonstrated based on the Example. It is to be understood by those skilled in the art that this embodiment is merely an example, and that various modifications are possible and that such modifications are within the scope of the present invention.

Claims (37)

An agent for promoting maintenance of undifferentiation of pigment stem cells, comprising an ATM or an ATM variant having an effect of promoting maintenance of differentiation of pigment cells. The ATM or the ATM variant is
(A) ATM consisting of the amino acid sequence represented by SEQ ID NO: 1,
(B) an ATM variant comprising an amino acid sequence having at least 80% homology with the amino acid sequence represented by SEQ ID NO: 1 and having an action to promote maintenance of undifferentiation of pigment stem cells;
(C) ATM encoded by the nucleotide sequence represented by SEQ ID NO: 2, or (d) Nucleic acid hybridized under stringent conditions to the nucleotide sequence represented by SEQ ID NO: 2 or its complementary strand, and a dye stem cell The agent for maintaining undifferentiation of pigment stem cells according to claim 1, which is any of the ATM variants having an action for promoting the maintenance of undifferentiation. A vector comprising a promoter sequence that controls so that ATM or a variant thereof is expressed in a region containing pigment stem cells, and a nucleotide sequence that encodes ATM or an ATM variant that is linked downstream of the promoter sequence. An agent for maintaining the undifferentiation of pigment stem cells. A nucleotide sequence encoding the ATM or the ATM variant is
(A) a nucleotide sequence encoding ATM consisting of the amino acid sequence represented by SEQ ID NO: 1,
(B) a nucleotide sequence encoding an ATM variant consisting of an amino acid sequence having at least 80% homology with the amino acid sequence represented by SEQ ID NO: 1 and having an action to promote maintenance of undifferentiation of pigment stem cells;
(C) hybridizes to the nucleotide sequence represented by SEQ ID NO: 2 or (d) the nucleotide sequence represented by SEQ ID NO: 2 or a complementary strand thereof under stringent conditions, and promotes the maintenance of undifferentiation of pigment stem cells A nucleotide sequence encoding an ATM variant having
The agent for maintaining undifferentiation of pigment stem cells according to claim 3, comprising a vector having any one of the following. An agent for maintaining undifferentiation of pigment stem cells, comprising an ATR variant having an ATR or pigment cell undifferentiation maintenance promoting action. The ATR or the ATR variant is
(A) ATR consisting of the amino acid sequence represented by SEQ ID NO: 3,
(B) an ATR variant comprising an amino acid sequence having at least 80% homology with the amino acid sequence represented by SEQ ID NO: 3, and having an action to promote maintenance of undifferentiation of pigment stem cells;
(C) ATR encoded by the nucleotide sequence represented by SEQ ID NO: 4, or (d) a nucleic acid that is hybridized under stringent conditions to the nucleotide sequence represented by SEQ ID NO: 4 or its complementary strand, and a dye stem cell The agent for promoting the maintenance of undifferentiation of pigment stem cells according to claim 5, which is any one of the ATR mutants having an action for promoting maintenance of undifferentiation. A vector comprising a promoter sequence that controls ATR or a variant thereof to be expressed in a region containing pigment stem cells, and a nucleotide sequence encoding ATR or an ATR variant linked downstream of the promoter sequence. An agent for maintaining the undifferentiation of pigment stem cells. A nucleotide sequence encoding the ATR or the ATR variant is
(A) a nucleotide sequence encoding ATR consisting of the amino acid sequence represented by SEQ ID NO: 3,
(B) a nucleotide sequence comprising an amino acid sequence having at least 80% homology with the amino acid sequence represented by SEQ ID NO: 3 and encoding an ATR variant having an action to promote maintenance of undifferentiation of pigment stem cells;
(C) hybridizes under stringent conditions to the nucleotide sequence represented by SEQ ID NO: 4 or (d) the nucleotide sequence represented by SEQ ID NO: 4 or a complementary strand thereof, and promotes the maintenance of undifferentiation of pigment stem cells A nucleotide sequence encoding an ATR variant having
The agent for maintaining undifferentiation of pigment stem cells according to claim 7, comprising a vector having any of the above. The agent for maintaining undifferentiation of pigment cells according to any one of claims 1 to 8, wherein the undifferentiation loss of the pigment stem cells is differentiation of the pigment stem cells into mature pigment cells in a bulge region. An agent for maintaining undifferentiation of pigment stem cells, comprising one or more proteins included in a protein group activated by a genome damage inducing agent. The agent for maintaining undifferentiation of pigment stem cells according to claim 10, comprising a regulatory factor that controls the activity of one or more proteins contained in the protein group activated by the genomic damage inducer. The agent for maintaining undifferentiation of pigment stem cells according to claim 11, wherein the regulator is any one of a compound, an oligonucleotide, a peptide, and a protein. A nucleotide sequence encoding one or more proteins included in a protein group activated by a genomic damage-inducing source, which is linked downstream of the promoter sequence and is controlled so as to be expressed in a region containing a pigment stem cell. Including a vector having
An agent for maintaining undifferentiation of pigment stem cells. An agent for maintaining undifferentiation of pigment stem cells, comprising a regulator that controls the activity of one or more proteins included in a group of proteins activated by a genome damage-inducing agent. The agent for maintaining undifferentiation of a pigment stem cell according to any one of claims 10 to 14, which suppresses differentiation of the pigment stem cell into a mature pigment cell in a bulge region caused by the genome damage inducer. An agent for promoting maintenance of self-renewal of pigment stem cells, comprising ATM or an ATM variant having an action of promoting self-renewal of pigment cells. A vector comprising a promoter sequence that controls so that ATM or a variant thereof is expressed in a region containing pigment stem cells, and a nucleotide sequence that encodes ATM or an ATM variant that is linked downstream of the promoter sequence. , An agent for maintaining the self-renewal of pigment stem cells. An agent for maintaining the self-renewal of pigment stem cells, comprising ATR or an ATR variant having an effect of promoting self-renewal of pigment cells. A vector comprising a promoter sequence that controls ATR or a variant thereof to be expressed in a region containing pigment stem cells, and a nucleotide sequence encoding ATR or an ATR variant linked downstream of the promoter sequence. , An agent for maintaining the self-renewal of pigment stem cells. An agent for maintaining self-replication of pigment stem cells, comprising one or more proteins included in a protein group activated by a genome damage-inducing agent. 21. The agent for maintaining the self-replication of pigment stem cells according to claim 20, comprising a regulatory factor that controls the activity of one or more proteins contained in the protein group activated by the genome damage-inducing agent. The control factor is any one of a compound, an oligonucleotide, a peptide, and a protein.
The agent for maintaining self-renewal of pigment stem cells according to claim 21. A nucleotide sequence encoding one or more proteins included in a protein group activated by a genomic damage-inducing source, which is linked downstream of the promoter sequence and is controlled so as to be expressed in a region containing a pigment stem cell. Including a vector having
Self-replication promoter for pigment stem cells. An agent for maintaining self-replication of pigment stem cells, comprising a regulatory factor that controls the activity of one or more proteins included in a protein group activated by a genome damage-inducing agent. 25. Inhibition of inhibition of self-replication of the pigment stem cells in a bulge region caused by abnormal differentiation of the pigment stem cells into mature pigment cells caused by the genomic damage-inducing source. The agent for maintaining self-renewal of the described dye stem cells. A mammalian depigmentation therapeutic agent comprising the pigment stem cell undifferentiation maintenance promoter according to any one of claims 1 to 15. A mammalian depigmentation therapeutic agent comprising the pigment stem cell self-replication inhibition inhibitor according to any one of claims 16 to 25. 27. The mammalian depigmentation is depigmentation by loss of the pigment stem cells.
Or the mammalian depigmenting therapeutic agent according to 27. An agent for maintaining undifferentiation of tissue stem cells, comprising one or more proteins included in a protein group activated by a genome damage inducing agent. 30. The tissue stem cell undifferentiation maintenance promoter according to claim 29, comprising a regulatory factor that controls the activity of one or more proteins contained in a protein group activated by the genome damage-inducing agent. The control factor is any one of a compound, an oligonucleotide, a peptide, and a protein.
The agent for maintaining undifferentiation of tissue stem cells according to claim 30. A promoter sequence that is controlled to be expressed in a region containing tissue stem cells;
Promoting maintenance of undifferentiation of tissue stem cells, comprising a vector having a nucleotide sequence encoding one or more proteins included in a group of proteins activated by a genome damage-inducing source, connected downstream of the promoter sequence Agent. An agent for maintaining undifferentiation of tissue stem cells, comprising a regulatory factor that controls the activity of one or more proteins included in a protein group activated by a genome damage-inducing agent. 34. The tissue stem cell undifferentiated maintenance promoter according to any one of claims 29 to 33, which suppresses differentiation of the tissue stem cell into a mature cell caused by the genome damage inducer. A therapeutic agent for suppressing aging in mammals, comprising the tissue stem cell undifferentiated maintenance promoter according to any one of claims 29 to 34. A cosmetic composition comprising one or more proteins included in a protein group activated by a genome damage inducer. A functional food comprising one or more proteins included in a protein group activated by a genome damage inducer.