KR20170124003A - Method for predicting skin pigmentation - Google Patents

Abstract

As a method for providing information about skin pigmentation, disclosed is a method for providing information about skin pigmentation, which comprises the following steps: measuring one or more of rs3846662 single nucleotide polymorphism (SNP) of a HMGCR gene, rs1799983 SNP of a NOS3 gene and rs9939609 SNP of a FTO gene from a specimen, and when the genotype of SNP position of the gene is E0, E1 and E2, the skin barrier function is excellent in order of E0>E1>E2. The genotype score S_G from each of the genes is calculated and analyzed.

Description

Method for predicting skin pigmentation "

This specification describes a method for predicting skin pigmentation.

Several factors are involved in determining the color of a person's skin. Among them, factors such as activity of melanocyte (melanocyte) which makes melanin pigment, distribution of blood vessels, thickness of skin, presence of pigment inside and outside the body such as carotinoid and bilirubin are important. In particular, the most important factor is melanin, a melanin pigment produced by the action of various enzymes such as tyrosinase in melanocytes in the human body. The formation of melanin pigment affects genetic factors, hormonal secretion, physiological factors such as stress, and environmental factors such as ultraviolet irradiation.

The melanin pigment produced in melanocytes of body skin is a phenolic polymer substance having a complex form of black pigment and protein. It protects the subcutaneous skin organs by blocking ultraviolet rays irradiated from the sun, and at the same time, free radicals And protects proteins and genes in the skin. Melanin, which is produced by stress stimuli inside and outside the skin, is a stable substance that does not disappear until the skin is excreted through skin keratinization even if the stress disappears. However, when melanin is produced more than necessary, it induces hypercholesterolemia such as spots, lobes, and dots, resulting in poor cosmetic results.

Today, oriental women prefer white, clean skin like white whites and use it as an important criterion of beauty, so there is a growing demand for improvement of hyperpigmentation and resolution of cosmetic problems.

However, due to the genetic and environmental influences, the individual skin characteristics are very different, so the effects of using the same method can vary widely. Therefore, it is important to understand individual skin characteristics in order to maintain and improve effective skin condition.

Genetic characteristics of individuals can be diagnosed through genes. For example, diagnostic methods using genetic markers are being conducted to identify the risk of developing a specific disease. In such a method, it is necessary to identify the marker having a relation with the target characteristic.

Furthermore, the skin characteristic is represented by a combination of various genetic characteristics, and there is a demand for a method for determining more accurate skin characteristic by revealing the factors involved in the characteristic.

Korean Patent Publication No. 10-2009-0027537 (Mar. 17, 2009)

In one aspect, a task to be solved by the present invention is to provide information about the ability of a person to genetically preserve pigmentation.

In another aspect, an object of the present invention is to accurately predict skin pigmentation of an individual through a specific genotype.

In another aspect, an object of the present invention is to provide an accurate prediction of an individual's inherent skin pigmentation prevention ability by reflecting the degree of association of a specific genotype and the degree of influence of a specific gene.

In another aspect, the object of the present invention is to provide a method of predicting an individual's ability to inhibit inherent skin pigmentation with high accuracy, taking into consideration the genetic characteristics of individual genes and the influence of specific genotypes and the influence of specific genes will be.

In one aspect of the present invention, rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism ) And rs16891982 single base polymorphism (SNP) of the SLC45A2 gene, and analyzing genetic pigment deposition according to the base at the SNP position of the gene.

In another aspect of the present invention, rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, and SLC45A2 A primer and a probe for detecting a base at one or more SNP positions of rs16891982 single nucleotide polymorphism (SNP) of the gene.

In another aspect of the present invention, there is provided a skin pigmentation prediction kit comprising the composition for predicting skin pigmentation.

In one aspect, the present invention can provide information about the ability of a person to genetically retain skin pigmentation.

In another aspect, the present invention can accurately predict skin pigmentation of an individual through a specific genotype.

In another aspect, the present invention reflects the degree of association of a particular genotype and the degree of influence of a particular gene, so that an individual can predict highly accurately the inherent skin pigmentation.

In another aspect, the present invention can accurately predict the inherent skin pigmentation prevention ability of an individual in consideration of the genetic characteristic of each race, taking into consideration the degree of association of a specific genotype and the influence of a specific gene.

In another aspect, the present invention provides a method for accurately predicting characteristics associated with skin pigmentation of an individual, and thus providing a management criterion for proper skin whitening.

FIG. 1 is a diagram illustrating a workflow of a single base polymorphism detection method according to the TaqMan probe method.
Figure 2 shows the nucleotide sequence representing the rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene.
Figure 3 shows the nucleotide sequence representing the rs1800414 single nucleotide polymorphism of the OCA2 gene.
Figure 4 shows the nucleotide sequence representing the rs885479 single nucleotide polymorphism of the MC1R gene.
Figure 5 shows the nucleotide sequence representing the rs1834640 single nucleotide polymorphism of the SLC24A5 gene.
6 shows the nucleotide sequence representing the rs16891982 single nucleotide polymorphism of the SLC45A2 gene.

Hereinafter, embodiments of the present invention will be described in more detail. However, the techniques disclosed in the present application are not limited to the embodiments described herein but may be embodied in other forms. It should be understood, however, that the embodiments disclosed herein are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit and scope of the invention.

In the present specification, " single nucleotide polymorphism " (SNP) refers to a base mutation in which a single nucleotide appears differently in DNA, and shows a deviation depending on an individual. Such single nucleotide polymorphism Protein structure, expression level, or expression pattern, and the like.

In the present specification, the rs number specifying the SNP is a reference number of NCBI (National Center for Biotechnology Information).

As used herein, the term " probe " refers to a polynucleotide, a variant thereof, or a polynucleotide having a base sequence complementary to a target site of a gene and a marker substance bound thereto.

As used herein, the term " primer " refers to a polynucleotide having a base sequence complementary to the end of a specific region of a gene used for amplifying a specific region corresponding to a target region of the gene using PCR Means a variant thereof. The primer does not need to be completely complementary to the end of a specific region and can be used if it is complementary enough to hybridize to the terminal to form a double-stranded structure.

As used herein, " hybridization " means that two single-stranded nucleic acids form a duplex structure by pairing complementary base sequences. Hybridization can occur not only in perfect complementarity between single-stranded nucleic acid sequences, but also in the presence of some mismatching nucleotides.

As used herein, the term " frequency of genotypes " refers to the frequency of occurrence of the genotype in the genetic pool as a percentage. The gene pool may be a pool of whole human species or a gene pool of a particular race or group with a common genetic association. For example, the frequency of genotypes in Asian gene pools can be found at http://asia.ensembl.org.

In one embodiment, the invention provides a method of providing information on skin pigmentation.

(SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene from the DNA of the sample, , Rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene and rs16891982 single nucleotide polymorphism (SNP) of the SLC45A2 gene, and analyzing genetic skin pigmentation inhibition ability according to the base at the SNP position of the gene can do.

The measurement of the single base polymorphism of the gene from the sample may include identifying a base at the SNP position of each gene from the DNA of the sample. The method for identifying a base is not particularly limited as long as it is a method used for detecting a single nucleotide polymorphism (SNP). For example, the nucleotide sequence may be analyzed by sequencing, , Or a method of hybridizing with sample DNA using a probe complementary to the above sequence to measure the degree of hybridization. For example, TaqMan ^TM using allele-specific hybridization  Probe method using allele-specific hybridization through polymerisation, restriction fragment length polymorphism (RFLP) method using restriction enzymes, melting temperature (Tm) A dynamic allele-specific hybridization (DASH) method, a pyrosequencing method using a polymerization reaction, a microarray method using an allele-specific extension, or the like can be used But is not limited thereto.

In one embodiment, measuring a single base polymorphism of a gene from the sample comprises: (a) obtaining DNA from the sample; (b) rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, And an rs16891982 single nucleotide polymorphism (SNP) of the SLC45A2 gene; And (c) identifying a base at the SNP position in the amplified DNA.

The specimen can be, but is not limited to, human epithelial cells, keratinocytes, fibroblasts or melanin-forming cells.

The human ASIP gene is a gene encoding the AGOUTI signaling protein, which is also referred to as an agouti gene. When the ASIP gene expression is increased, the production of melanin melanin is decreased and the production of yellow melanin is increased. Depending on the rs6058017 SNP of the ASIP gene, there may be a difference in the synthesis of hair, skin and eye pigment.

The polynucleotide corresponding to rs6058017, which is the single nucleotide polymorphism (SNP) of the ASIP gene, has a single base polymorphism in which the base is G or A at the SNP position. The genotype of the single nucleotide polymorphic position may be GG, GA or AA.

In the ASIP gene, when the genotype of the SNP locus is GG, GA or AA, it can be determined that the genetic skin pigmentation prevention ability is superior in order of GG> GA> AA.

The human OCA2 gene encodes melanocyte-specific transporter protein located on the surface of melanocytes, and plays a role of transporting tyrosine, a precursor of melanin, into cells. A difference in melanin production may occur depending on the rs1800414 SNP of OCA2.

The polynucleotide corresponding to rs1800414, the single nucleotide polymorphism (SNP) of the OCA2 gene, has a single base polymorphism in which the base is G or A at the SNP position. The genotype of the single nucleotide polymorphic position may be GG, GA or AA.

In the OCA2 gene, when the genotype of the SNP locating base is GG, GA or AA, it can be judged that the OCG2 gene is genetically superior in skin pigmentation prevention ability in the order of GG> GA> AA.

The human MC1R gene is a gene encoding a melanocyte stimulating hormone receptor. When the function of the receptor is decreased, the production of melanin involved in black or black-brown in the cell is reduced. The rs885479 SNP of the MC1R may affect the production of black or dark brown melanin.

The polynucleotide corresponding to rs885479, which is the single nucleotide polymorphism (SNP) of the MC1R gene, has a single base polymorphism in which the base is A or G at the SNP position. The genotype of the single nucleotide polymorphic position may be AA, AG, or GG.

In the MC1R gene, when the genotype of the SNP locating base is AA, AG or GG, it can be judged that the genetic skin pigmentation prevention ability is superior in order of AA> AG> GG.

The human SLC24A5 gene encodes a potassium ion-dependent ion transport protein, which is located in the Golgi complex of melanocytes and plays an important role in the melanin formation process. The rs1834640 SNP of SLC24A5 may affect melanin production.

The polynucleotide corresponding to rs1834640, which is the single nucleotide polymorphism (SNP) of the SLC24A5 gene, has a single base polymorphism with a base of A or G at the SNP position. The genotype of the single nucleotide polymorphic position may be AA, AG, or GG.

In the SLC24A5 gene, when the genotype of the SNP locus is AA, AG or GG, it can be judged that the genetic skin pigmentation prevention ability is superior in order of AA> AG> GG.

The human SLC45A2 gene is a gene encoding a transport protein that mediates melanin synthesis and contributes to light skin. The rs16891982 SNP of SLC45A2 can affect melanin synthesis in skin, hair and eyes.

The polynucleotide corresponding to rs16891982, which is the single nucleotide polymorphism (SNP) of SLC45A2, has a single base polymorphism in which the base is C or G at the SNP position. The genotype of the single nucleotide polymorphic position may be CC, CG, or GG.

In the SLC45A2 gene, when the genotype of the SNP locus is CC, CG or GG, it can be judged that the genetic skin pigmentation prevention ability is superior in order of CC> CG> GG.

In one embodiment, rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene ) And the analysis of two or more of the rs16891982 single nucleotide polymorphisms (SNPs) of the SLC45A2 gene can be combined to determine genetic skin pigmentation deficits. It is possible to provide more accurate information about skin pigmentation by adjusting the contribution of each gene according to the degree of SNP of each gene having different function to skin pigment.

In another embodiment, the method of providing information comprises obtaining rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 One or more of the single nucleotide polymorphism (SNP) and the rs16891982 single nucleotide polymorphism (SNP) of the SLC45A2 gene were measured and the genotype score S _G And analyzing it.

The genetic skin pigmentation prevention ability of the skin can be determined according to the base at the SNP position of the gene. By confirming the SNP sequence of the gene, individuals can confirm functions such as brightness, melanin production, and specific melanin production of genetically innate skin. By checking the SNP sequence of the gene, an individual can determine genetically innate skin pigmentation prevention ability.

When genotypes of SNP loci of the genes are E0, E1 and E2, the ability to prevent skin pigmentation is excellent in order of E0 > E1 > E2, and scores can be given according to the genotypes.

The genotype score S _G S _G = 1 if the genotype of the specimen is E0, 0.5 ≤ S _G ≤ 0.8 if E1, and 0.2 ≤ S _G ≤ 0.5 if the frequency F E0 of the genotype of _E0 is less than 45%. For example, S _G = 1 for E 0, 0.55 ≤ S _G ≤ 0.75 for E 1, 0.25 ≤ S _G ≤ 0.45 for E 2, and S _G = 1 for E 0 and 0.6 ≤ S _G ≤ 0.7 for E 1, , Then 0.3 < = S _G < / = 0.4. However, the genotype scores of E0, E1 and E2 are E0>E1> E2. In one example, the genotypic score was a frequency of E0 genotype, rs6058017 single nucleotide polymorphism (SNP) of ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of MC1R gene, rs1834640 single gene of SLC24A5 gene (SNP) and the genotype score for the rs16891982 single nucleotide polymorphism (SNP) of the SLC45A2 gene. In one example, the genotypic score may be appropriately adjusted in consideration of the genotype ratio appearing in the sample pool.

The genotype score S _G can be represented by the following formula 1 when the frequency F of the _E0 E0 genotype is not less than 45%.

<Formula 1>

Genotype score (S _G ) = F _s / F _E0

F _s is the frequency of the genotype of the specimen, and F _E0 is the frequency of the E0 genotype.

Since the genotype score is calculated according to the frequency of the specific genotype as described above and the frequency in the actual gene pool is reflected, more matching information can be provided for the specimen. In particular, it is possible to provide more accurate information by limiting the gene pool to a specific racial or genetic association group to which the specimen belongs.

The genotypes E0, E1 and E2 described above are described according to the order of inhibiting skin pigment deposition according to the SNP base genotype of each of the genes described above, and specifically shown in Table 1 below.

Gene name Reference SNP genotype E0 E1 E2 ASIP rs6058017 GG GA AA OCA2 rs1800414 GG GA AA MC1R rs885479 AA AG GG SLC24A5 rs1834640 AA AG GG SLC45A2 rs16891982 CC CG GG

When the genotypic score for two or more SNPs of the gene is reflected in the provision of the information, the genotypic score for each gene may be determined according to the degree of relevance of each gene to the skin pigment in the genetically relevant group to which the subject belongs You can adjust the percentage of points to be reflected.

In one example, the genotype score for the rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene can be reflected at a rate of 10-50%, 20-40%, or 25-35%.

In one example, the genotype score for the rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene may be reflected at a rate of 5-40%, 10-30%, or 15-25%.

In one example, the genotype score for the rs885479 single nucleotide polymorphism (SNP) of the MC1R gene can be reflected at a rate of 10-50%, 20-40%, or 25-35%.

In one example, the genotype score for the rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene may be reflected at a rate of 0.5-30%, 1-20%, or 5-15%.

In one example, the genotype score for the rs16891982 single nucleotide polymorphism (SNP) of the SLC45A2 gene may be reflected at a rate of 0.5-30%, 1-20%, or 5-15%.

For example, rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, and SLC45A2 gene Rs16891982 genotype score for single nucleotide polymorphism (SNP) S _G Can be reflected at a ratio of 1-5: 0.5-4: 1-5: 0.1-3: 0.1-3, for example, 2-4: 1-3: 2-4: 0.1-2: 0.1-2 Of the total. When the ratio is reflected, it is possible to provide more accurate information about genetic skin pigment.

The information providing method according to the embodiments of the present invention can provide more accurate information by not only determining the single nucleotide polymorphisms related to the five genes but also calculating the genotype score reflecting the frequency of the genotypes. In addition, it is possible to provide more accurate information by reflecting each gene at a specific rate, rather than simply combining the judgments about these genes.

In one embodiment, the skin pigment score can be calculated reflecting the genotype score for the five genes. The rate of reflection of the genotypic score may be as described above. According to the score, the classification regarding skin pigmentation can be classified to provide information on the skin pigment. In one example, the grade may be classified into good, interest, and attention in order of excellent skin pigmentation prevention ability, but not limited thereto.

The rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, the rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, the rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, the rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, And the nucleotide sequence of rs16891982 single nucleotide polymorphism (SNP) of the SLC45A2 gene. The SNP base sequence of the ASIP gene is SEQ ID NO: 1, the OCP2 gene SNP base sequence is SEQ ID NO: 2, the MC1R gene SNP base sequence is SEQ ID NO: 3, the SLC24A5 gene SNP base sequence is SEQ ID NO: 4, Is represented by SEQ ID NO: 5. The bases in brackets in Figs. 2-6 represent bases at the SNP position.

In one embodiment, one or more of a primer and a probe may be used to detect a base at the SNP position of the gene.

The probe may comprise 20 to 60, for example 40 to 52, for example 45 to 50 consecutive nucleotide sequences comprising a base at the SNP position of the nucleotide sequence, or a sequence complementary to the nucleotide sequence . The probe may have a sequence that is capable of hybridizing to the SNP sequence to detect a base at the SNP position. The probe may comprise a polynucleotide having such a sequence and a labeling substance bound thereto for detection.

The primer may comprise a set of primers consisting of two polynucleotides having a sequence complementary to each of 10 to 40 consecutive nucleotide sequences at the 5 'and 3' ends of the gene fragment of the target site. The primer set is used for amplification of a target site, and the target site is a contiguous base sequence comprising a base at the SNP position of the gene, for example, 50 to 400 consecutive bases containing a base at the SNP position Base sequence. The target site may be, for example, 100 to 300 consecutive nucleotide sequences, including, for example, 200 consecutive nucleotide sequences containing a nucleotide at the SNP position.

In one embodiment of the invention, rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 SNP of the SLC24A5 gene, And a primer and a probe for detecting a base at one or more SNP positions in the rs16891982 single nucleotide polymorphism (SNP) of the SLC45A2 gene, and a probe for predicting skin pigmentation. The primer and the probe are the same as those described for the primer and the probe for detecting the base at the SNP position of the above-mentioned gene, and the description is omitted.

In one embodiment of the present invention, a skin pigmentation prediction kit comprising the skin pigment deposition prediction composition can be provided.

According to embodiments of the present invention, information about skin pigmentation that an individual has genetically possessed can be provided. Specifically, it is possible to identify a single base polymorphism of a specific gene involved in genetic skin pigmentation, thereby providing information on an individual's natural skin characteristics, thereby providing appropriate information on management, treatment, maintenance, etc. Can be provided.

Hereinafter, the present invention will be described in detail with reference to Examples, Comparative Examples and Test Examples. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not to be construed as limiting the scope of the present invention. It will be self-evident.

[Test Example 1] DNA extraction and amplification from a specimen

Genomic DNA was extracted from blood of subjects using blood DNA (QIAamp DNA Blood Mini Kit, QIAGEN) in 108 Koreans. The extracted DNA was purified and used in the following experiments.

[Test Example 2] Detection of gene SNP

Using the DNA of the subject amplified and purified in Test Example 1 as a template, the SNP of each gene was detected by the TaqMan probe method as shown in Fig.

Specifically, 12.5 .mu.l of 2X TaqMan Universal PCR Master mix, 20 .mu.l Primer and TaqMan Probe dye mix of TaqMan SNP Genotyping Assay specific to each gene SNP according to the following Table 2 was added to 20ng of the subject DNA obtained in Test Example 1 , And 11.25 μl of DNase-free sterile water. The final reaction solution was adjusted to 25 μl by vortexing. The reaction solution was dispensed into a 96-well plate to perform PCR reaction. The PCR conditions were 95 ℃ for 10 min, followed by denaturation at 95 ℃ for 15 sec and annealing and extension at 60 ℃ for 40 cycles. After that, gene SNP was detected using a real-time PCR system (TaqMan Mastermix, ABI).

Gene name Reference SNP TaqMan SNP Genotyping Assay ID ASIP rs6058017 C__22275334_10 OCA2 rs1800414 C___8866240_10 MC1R rs885479 C___7519060_20 SLC24A5 rs1834640 C___2908200_10 SLC45A2 rs16891982 C___2842665_10

 [Test Example 3] Genotype analysis and genotype score calculation

Using the SNP genotype identified in Test Example 2, the genotype score S _G Respectively.

Gene name Reference SNP genotype E0 E1 E2 ASIP rs6058017 GG GA AA OCA2 rs1800414 GG GA AA MC1R rs885479 AA AG GG SLC24A5 rs1834640 AA AG GG SLC45A2 rs16891982 CC CG GG

The frequency (F _E0 , F _E1 , F _E2 ) for each of the SNP genotypes in Table 3 is obtained from http://asia.ensembl.org and when the frequency F E0 of the genotype of _E0 is less than 45% And the frequency of the genotype of _E0 was 45% or more, it was determined by the following equation (1).

<Formula 1>

Genotype score (S _G ) = F _s / F _E0

The genotypic score of each gene was reflected at the ratio of 3: 2: 3: 1: 1 in the order of Table 3, and the scores for skin pigmentation were shown in Table 4 below.

gene Reference SNP E0 E0 Score E1 E1 Score E2 E2 Score ASIP rs6058017 GG 30.0 AG 20.0 AA 10.0 OCA2 rs1800414 GG 20.0 GA 10.0 AA 5.0 MC1R rs885479 AA 30.0 AG 20.0 GG 10.0 SLC24A5 rs1834640 AA 10.0 AG 9.0 GG 8.0 SLC45A2 rs16891982 CC 10.0 CG 9.0 GG 8.0

[Test Example 4] Evaluation of skin pigmentation

Skin pigmentation of the subject was evaluated by the following method.

[Test Example 5] Confirmation of correlation between skin pigmentation score and skin pigmentation

The correlation between the skin pigmentation score calculated in Test Example 3 and the skin pigmentation measured in Test Example 4 was analyzed.

From the above results, it can be seen that information on skin pigmentation and whitening can be more accurately provided to an individual by discriminating the skin pigmentation through the genotype score in consideration of the genotype frequency of the specific gene SNP of the present invention.

<110> AMOREPACIFIC CORPORATION <120> Method for predicting skin pigmentation <130> 16P194 / IND <160> 5 <170> KoPatentin 3.0 <210> 1 <211> 1001 <212> DNA <213> Homo sapiens <400> 1 ccactgcagt ccagcctggc tgacagagtg aaaccctgtc taaaaaaaaa acaaaaaaca 60 aacaaacaaa aaaaacacaa actagggaga agacgaccag ggccaggcaa gagccaggta 120 agtctggatg gggatggagg tgagggaatc tgactggggg agcgggcggt gggcggtggg 180 caagccagcg gggaaacctc tggtccggga tctccctagc ccgaggagct cccaggcaga 240 ggtgaggacc ggaggggtgg gcgtggccgg ctcataaagc cccggcgttt cccacgcaga 300 aggaggcttc gatgaagaaa gtggtgcggc cccggacccc cctatctgcg ccctgcgtgg 360 ccacccgcaa cagctgcaag ccgccggcac ccgcctgctg cgacccgtgc gcctcctgcc 420 agtgccgctt cttccgcagc gcctgctcct gccgcgtgct cagcctcaac tgctgagcgc 480 ccccactccc ggccgcgagc rggcagggct tcggggacgc ggggcgcttc tcgggcgggt 540 gatccctaac agggcggctt cccagggctg caggcgggcg gaggttccag gagatgggac 600 ttcagggaga cctggcttgg gctaaaatcg aaatacaata tatataggct gctcgaaggt 660 gtgcggctgt ttctgtaaag gtcccgaaag ttctcggtcc cttcgccacg gagtcccgcg 720 ctgctctccc ggggctcagg aaacccgggc gtcccagcct cacagctgca ctcccagtca 780 ccgctgctgc cgacctgggg gttaggaacg aggctatccc ctgcctttaa acctgggaac 840 acccgaggct gcctccaaaa ccaggaaatt cagaccagcc aggaagaaag gaacagcctt 900 tcagtctcac acatttcctc ctaggtgctc ccctctcccc aggggaaaaa aaaaacggaa 960 agaaagaaag aggccaagcg cggtggctca cgcctgtaat c 1001 <210> 2 <211> 1001 <212> DNA <213> Homo sapiens <400> 2 ccctccccct atgaccaggt ggattctcgc gggtgtcatt ccagcgctaa gagtgcaccc 60 cctgcattcc aggggcctca tgcatgtcgt gtaaagaaca tggccagagc cttatctggg 120 agtcacagtt ccgtgagaag gctcagcctc atggccccac ccgcatgctt ggcgtggtag 180 agaaaggaac agtgaagaca gcataggccc ctgtagaaac gccccgtcat cctctgatac 240 ctgccggcca ggtgtttcat aacagggctg tgctactctt gacatctgtg tttatctttc 300 ataaagattt tgaatgcagt aatcctgaat ctgtacgggt ttccttgtaa cacagtactt 360 tgccattttc tttcaagttc gagaggttac atttttcatc ctcgtgaaat ctgtcgtgat 420 tccagttgcg taggttatga cacgctgcag gagtcagaag gttgtgcaga gtaaatgagc 480 tgtggtttct ctcttacagc rtaggatatc tgacgggatt ctgctcgcca aatgcctgac 540 agtgttggga tttgttatct tcatgttttt cctcaattcg tttgtccctg gcattcatct 600 tgatcttggt gagtctaatt tagctttggt tcataggctt tgtcacattc tggatgggaa 660 ggtttcagag cctgttccca gacactgact ttgcccacag gcagccgggc tggtggaagg 720 ccagagaggg ctgagatgga gggtgggcag cctgccctgg gaagaagggc gcctttcctt 780 ttggtttcct gggcaggagg gagggagaga gagatgcatc tctggcccct tagactctgt 840 gccatgggtc ctcagcccct ccagggatga ccatgaggag gaatatagag tgggcactgt 900 cctgtctatt gtagttaata accacatctt tacatggttc ccagaagaga tggagccaca 960 tgggcaaggc cagcgctgcc atctgtgccg cctaccatgc c 1001 <210> 3 <211> 1001 <212> DNA <213> Homo sapiens <400> 3 cctggacagg actatggctg tgcagggatc ccagagaaga cttctgggct ccctcaactc 60 cacccccaca gccatccccc agctggggct ggctgccaac cagacaggag cccggtgcct 120 ggaggtgtcc atctctgacg ggctcttcct cagcctgggg ctggtgagct tggtggagaa 180 cgcgctggtg gtggccacca tcgccaagaa ccggaacctg cactcaccca tgtactgctt 240 catctgctgc ctggccttgt cggacctgct ggtgagcggg agcaacgtgc tggagacggc 300 cgtcatcctc ctgctggagg ccggtgcact ggtggcccgg gctgcggtgc tgcagcagct 360 ggacaatgtc attgacgtga tcacctgcag ctccatgctg tccagcctct gcttcctggg 420 cgccatcgcc gtggaccgct acatctccat cttctacgca ctgcgctacc acagcatcgt 480 gaccctgccg cgggcgcggc ragccgttgc ggccatctgg gtggccagtg tcgtcttcag 540 cacgctcttc atcgcctact acgaccacgt ggccgtcctg ctgtgcctcg tggtcttctt 600 cctggctatg ctggtgctca tggccgtgct gtacgtccac atgctggccc gggcctgcca 660 gcacgcccag ggcatcgccc ggctccacaa gaggcagcgc ccggtccacc agggctttgg 720 ccttaaaggc gctgtcaccc tcaccatcct gctgggcatt ttcttcctct gctggggccc 780 cttcttcctg catctcacac tcatcgtcct ctgccccgag caccccacgt gcggctgcat 840 cttcaagaac ttcaacctct ttctcgccct catcatctgc aatgccatca tcgaccccct 900 catctacgcc ttccacagcc aggagctccg caggacgctc aaggaggtgc tgacatgctc 960 ctggtgagcg cggtgcacgc ggctttaagt gtgctgggca g 1001 <210> 4 <211> 1001 <212> DNA <213> Homo sapiens <400> 4 ttaagatgtg atggaaaaat atggcagatt gtgttgcctg tggtatatct taagaatcaa 60 cttcagaggc tttcatttca atacttcatc aaaaatagac atctggataa ccccttctcc 120 ttctgcctac ccattattta aaactattct acaaatattc tacctgggaa taaactcctc 180 cttctttaag tgcccacaat gccttgtaac ttgagtctag cccctgttaa ttcctgcctt 240 attttaagtt tattcacatg ttatagttgt tgttgactat gttgcaactt ttgcactggg 300 ccagcagaaa cctgaggtca ggtctcctgc ctttatttct catgctcagt ttggaatcca 360 tgttcatttt cgcgttgtgt catccttgtg gctccagcct gcattaatag ccccctttgc 420 tgcatgagca acaaccgtta gagacccata cttgtagtga ctgagacaca gtgacattat 480 atcacaacct cagaaaccac racataaacc aaggaataat caatgccata gtttttaata 540 gtgcaactag aaatggattg tctgctgcta agtataggga aacagttata gctggagctg 600 ctgcagaggg gagggatgat gtttaataat gtgatatagg aaatctcacg atgctacaga 660 aattgattat ctattcttag cttcaacctg catctgactt ttcccacata aagggggaat 720 aaactccatc aaaaatcaga gcccttaaaa tatcagcttc tggccctcaa attactaatc 780 ctcacagagg aaaggcagct ctccaatcac tctcttctta gagccacaaa actcagcgat 840 ctacatgacc ataggcagca tgatggaggg atactgaagg gaaaccaggt cttctaaaga 900 atgtaactga tggacccagg gccactgaca tcagagccag aaacattgtt ttctttttaa 960 caacttaaat ctgtctaacc tctcatgtta ttggggcctg g 1001 <210> 5 <211> 1001 <212> DNA <213> Homo sapiens <400> 5 ctcctataac ccacccagtt acacatctga gctaaggtca ttttgcatgc tgactttgtc 60 acctgtcata ttaatattgc ttctcttctg tatcatctac aaatctgata agcctatcta 120 tgaaatgcta ccaatgatct tgctccagta gaacacgaac ctgctgggac tcatccatcc 180 aacacttcat cttgtaaaga ttccctttca ttttccagag aaacttgatc aggaacccac 240 tgattccaag agcaaagtaa tcagtgagga aatgacacct agaattcatg atgaaaaaag 300 gatgctttat atggtccttt ttaaggtgat agtttttcct gacgtccata gatttattaa 360 gaatctggta ttttaaacag taggaaatac acatagaaat atcaaatcca agttgtgcta 420 gaccagaaac ttttagaaga catccttagg agagagaaag acttacaaga ataaagtgag 480 gaaaacacgg agttgatgca saagccccaa catccaacct cgactcctct ttcgtagatg 540 agaaactctg tggagttgtg tgcactatag ggatccccgc ggtacacaat ctgaaagaga 600 gattggaggc tgttgaggta caaatgcaat gtagtaagaa cccttctcat gcactctgct 660 tctccacctc tggggagcaa atgtccctgc ctgagggacc ctttctagag tacagagctg 720 atgctgtcat gaccaagtcg catcctatca catcttcatt caaggaccta tcagaaatct 780 aatgcttgct gcatcagatc tcaacaacac tctcttggat ccttaacatt cctccttcca 840 tctcacagct ggccccacct gacttaggaa atcccaccac tcatcttctg ctacagccaa 900 tcggcttctg actcatctcc acatgggcca ttcctgttcc tgcttggaat gttttctccc 960 tctctaatat tcctctaact cctactcacc cttcaaggcc c 1001

Claims (26)

As a method for providing information on skin pigmentation,
The rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, and rs16891982 Measuring at least one of a single nucleotide polymorphism (SNP), and analyzing genetic pigmentation according to a base at a SNP position of the gene. The information providing method according to claim 1,
(a) obtaining DNA from a specimen;
(b) rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, And an rs16891982 single nucleotide polymorphism (SNP) of the SLC45A2 gene; And
(c) analyzing the skin pigmentation according to the base at the SNP position by identifying a base at the SNP position in the amplified DNA. The method according to claim 1,
Wherein said ASIP gene is judged to be excellent in genetic skin pigmentation prevention ability in the order of GG>GA> AA when the genotype of said SNP locating base is GG, GA or AA. The method according to claim 1,
Wherein said OCA2 gene is judged to be superior in genetic skin pigmentation prevention capability in the order of GG>GA> AA when the genotype of said SNP locus is GG, GA or AA. The method according to claim 1,
Wherein said MC1R gene is judged to be superior in genetic skin pigmentation prevention ability in the order of AA>AG> GG when the genotype of the SNP locating base is AA, AG or GG. The method according to claim 1,
Wherein the genetic type of the SNP locus in the SLC24A5 gene is AA, AG, or GG, and determining that the genetic skin pigmentation prevention capability is superior in order of AA>AG> GG. The method according to claim 1,
Determining that the SLC45A2 gene is superior in genetic skin pigmentation prevention capability in the order of CC>CG> GG when the genotype of the SNP locating base is CC, CG or GG. The method according to claim 1,
The rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, the rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, the rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, the rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, and the SLC45A2 gene One or more of the rs16891982 single nucleotide polymorphisms (SNPs) were measured,
When genotypes of SNP loci of the genes are E0, E1 and E2, the genetic skin pigmentation prevention ability is excellent in order of E0 > E1 > E2,
From each of the genes, the genotype score S _G And analyzing it,
The genotype score S _G The frequency of the genotype of _E0 is less than 45%. If the genotype of the specimen is E0, S _G = 1, if E0 is 0.5 ≤ S _G ≤ 0.8, and if E2 is 0.2 ≤ S _G ≤ 0.5, _{And E0} is 45% or more,
How to provide information:
<Formula 1>
Genotype score (S _G ) = F _s / F _E0
F _s is the frequency of the genotype of the specimen, F _E0 is the frequency of the E0 genotype, and F _s and F _E0 are obtained from the genotype database of the race to which the specimen belongs. 9. The method of claim 8,
The rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, and rs16891982 single Reflecting the genotype score S _G for two or more of the base polymorphisms (SNPs)
And reflecting the genotype score of rs6058017 single nucleotide polymorphism (SNP) of said ASIP gene at a rate of 10 to 50%. 9. The method of claim 8,
The rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, and rs16891982 single Reflecting the genotype score S _G for two or more of the base polymorphisms (SNPs)
And reflecting the genotypic score for the rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene at a rate of 5 to 40%. 9. The method of claim 8,
The rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, and rs16891982 single Reflecting the genotype score S _G for two or more of the base polymorphisms (SNPs)
And reflecting the genotype score of rs885479 single nucleotide polymorphism (SNP) of the MC1R gene at a rate of 10 to 50%. 9. The method of claim 8,
Rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, and rs16891982 single Reflecting the genotype score S _G for two or more of the base polymorphisms (SNPs)
And reflecting the genotypic score of the SLC24A5 gene to rs1834640 single nucleotide polymorphism (SNP) at a rate of 0.5 to 30%. 9. The method of claim 8,
The rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, and rs16891982 single Reflecting the genotype score S _G for two or more of the base polymorphisms (SNPs)
And reflecting the genotype score of rs16891982 single nucleotide polymorphism (SNP) of the SLC45A2 gene at a rate of 0.5 to 30%. 9. The method of claim 8,
The rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, and rs16891982 single The genotype score for base polymorphism (SNP) S _G In a ratio of 1-5: 0.5-4: 1-5: 0.1-3: 0.1-3. 9. The method of claim 8,
Wherein the genotype of the SNP locating base of the ASIP gene is GG, GA or AA, and GG > GA > AA is superior in genetic skin pigmentation prevention capability. 9. The method of claim 8,
Wherein the genotype of the SNP locus of the OCA2 gene is GG, GA or AA, and GG > GA > AA is superior in genetic skin pigmentation prevention capability. 9. The method of claim 8,
Wherein the genotype of the SNP locus of the MC1R gene is AA, AG, or GG, and AA > AG > GG in the order of genetic skin pigmentation prevention. 9. The method of claim 8,
Wherein the genotype of the SNP locating base of the SLC24A5 gene is AA, AG or GG, and AA > AG > GG in the order of genetic skin pigmentation prevention. 9. The method of claim 8,
Wherein the genotype of the SNP locating base of the SLC45A2 gene is CC, CG or GG, and CC>CG> GG is superior in genetic skin pigmentation prevention capability. 9. The method of claim 8,
The genotype score S _G S _G = 1 if the genotype of the specimen is E0, 0.6? S _G ? 0.7 if E1, 0.3? S _G ? 0.4 if the frequency F E0 of the genotype of _E0 is less than 45%. 15. The method of claim 14,
The rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, and rs16891982 single The genotype score for base polymorphism (SNP) S _G At a ratio of 2-4: 1-3: 2-4: 0.1-2: 0.1-2,
The genotype score S _G S _G = 1 if the genotype of the specimen is E0, 0.55? S _G ? 0.75 if E1, 0.25? S _G ? 0.45 when _E0 is less than 45%. 22. The method according to any one of claims 1 to 21,
Wherein the sample is an epithelial cell, a keratinocyte, a fibroblast, or a melanin-forming cell isolated from a human. Rs6058017 single nucleotide polymorphism (SNP) of the ASIP gene, rs1800414 single nucleotide polymorphism (SNP) of the OCA2 gene, rs885479 single nucleotide polymorphism (SNP) of the MC1R gene, rs1834640 single nucleotide polymorphism (SNP) of the SLC24A5 gene, and rs16891982 single nucleotide polymorphism of the SLC45A2 gene A composition for predicting skin pigmentation comprising at least one of a primer and a probe for detecting a base at one or more SNP positions in a polymorphism (SNP). 24. The method of claim 23,
Wherein the probe is a polynucleotide having a sequence complementary to 20 to 60 consecutive nucleotide sequences comprising a base at the SNP position in the nucleotide sequence of the gene. 24. The method of claim 23,
Wherein the primer comprises a set of primers consisting of two polynucleotides having a sequence complementary to each of the consecutive 10 to 40 nucleotide sequences at the 5 ' end and the 3 ' end of the gene fragment, , Wherein said target site is a consecutive 50 to 400 nucleotide sequences comprising a base at said SNP position in the nucleotide sequence of said gene. 25. A skin pigmentation prediction kit comprising the composition for predicting skin pigmentation according to any one of claims 23 to 25.

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