EP0988399A1 - Dna sequences rich in triplet repeat useful in the diagnosis of trinucleotide repeat disorders - Google Patents

Dna sequences rich in triplet repeat useful in the diagnosis of trinucleotide repeat disorders

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Publication number
EP0988399A1
EP0988399A1 EP98929536A EP98929536A EP0988399A1 EP 0988399 A1 EP0988399 A1 EP 0988399A1 EP 98929536 A EP98929536 A EP 98929536A EP 98929536 A EP98929536 A EP 98929536A EP 0988399 A1 EP0988399 A1 EP 0988399A1
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Prior art keywords
sequence
sequences
seq
type
cdna
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German (de)
French (fr)
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Christian Neri
Howard M. Cann
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Fondation Jean Dausset-CEPH
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Fondation Jean Dausset-CEPH
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Definitions

  • the present invention relates to the detection of human genes comprising sequences with repeated triplets CAG / CTG or CGG / GCC, as well as the use of these genes in the diagnosis and the possible treatment of certain monogenic diseases or polygenic diseases with component hereditary.
  • the abnormal polymorphism of CAG / CTG or CGG / GCC repeating triplet sequences is a mutation involved in at least ten inherited human diseases (called repeating triplet diseases) that are primarily, but not exclusively, neurodegenerative diseases.
  • the expansion (several supernumerary triplets in patients) of repeated CAG / CTG triplet sequences is involved in particular in spinobulbar atrophy (SBMA), myotonic dystrophy (DM), spinocerebellar ataxia (SCA1) SCA1, SCA2, SCA3 , and SCA6, dendato-rubro-pallydoluysiane atrophy (DRPLA), and Huntington's disease (1 to 3).
  • SBMA spinobulbar atrophy
  • DM myotonic dystrophy
  • SCA1 spinocerebellar ataxia
  • SCA1 dendato-rubro-pallydoluysiane atrophy
  • DRPLA dendato-rubr
  • CGG / GCC repeat triplet sequences are notably implicated in fragile X syndrome (1).
  • restricted polymorphism of sequences with a repeated CAG / CTG triplet is notably implicated in Behçet's disease, an autoimmune disease for which the passage of 5 to 6 GCT codons coding for a polyalanine is associated with the disease (4).
  • CAG / CTG or CGG / GCC also called dynamic mutation
  • the expansion of CAG / CTG or CGG / GCC is often associated with a phenomenon of anticipation, the size of the repetitions often being inversely correlated with the age of onset and / or the severity of symptoms.
  • the expansion of the CAG / CTG repeats can appear in the non-coding (DM) or coding regions (for example SBMA, HD, DRPLA, SCAs) of the transcripts. These transcripts are sometimes expressed in tissues other than the brain and when they are translated lead to products of larger genes and carrying an abnormally elongated polyglutamine domain (1 to 3).
  • the expansion of repeated CAG / CTG triplets can be very large (for example DM; several hundred triplets), moderate (for example HD or SCA1; in general more than 35 triplets), or even very moderate (for example SCA6; in general less than 35 triples). In the normal state, these repetitions can be highly polymorphic
  • a variation of a single triplet within a short, weakly polymo ⁇ hical repeated triplet can also cause disease (4).
  • Autism and familial dementia which show variability in the age of onset or in the severity of symptoms, could also be caused by dynamic mutations.
  • a fairly large number of diseases could also originate or involve dynamic mutations: Parkinson's disease, spastic paraplegias, cerebrospinal ataxias not previously listed, - zonular cataracts, uncontrollable tremors, familial amyloid neuropathies, granulomatous arthritis familial (Blau's disease), hemifacial microsomies, - certain anemias and glaucomas, obsessive-compulsive disorders. • The above demonstrates the considerable importance of repeated triplets for the diagnosis and treatment of several diseases.
  • the present invention is based on a systematic study of the CAG / CTG or CGG / GCC repeats (which will be designated below by "[CAG] n" and “[CGG] n” respectively, n being the number of repetitions of the triplet), this study having been carried out with human cDNA libraries, these having undergone a first general screening, then the selected sequences having been selected on the basis of a certain number of criteria making it possible to ensure that the sequences in question belonged to new human genes and could, with high probability, be implicated in repeated triplet diseases. Finally, the selected sequences were the subject of a more complete study intended to allow their localization.
  • cDNA from human brains were studied to analyze the presence of CAG repeats using the hybridization of oligonucleotides on high density membranes.
  • the two libraries were obtained from mRNAs using the oligo-dT primer, one of the libraries being made up of fetal brain (FB) cDNA clones and the other library being made up of clones of Normalized newborn brain cDNA (NIB).
  • FB fetal brain
  • NBI Normalized newborn brain cDNA
  • the present invention is based on the demonstration of certain sequences capable of being involved in repeated triplet diseases obtained under hybridization conditions making it possible to select cDNAs which contain a number of [CAG] or [ CGG] greater than 7, which are more likely to be polymorphous in a sick population.
  • the complete conditions for analysis and selection of these different sequences will not be detailed below, only the elements making it possible to locate and re-isolate them will be provided, in particular, using the PCR primers which will be described below.
  • the present invention relates to a transcribed DNA sequence rich in repetitive triplet CAG / CTG or CGG / GCC corresponding to sequences A to L of the table, as well as their normal and mutated alleles and the complementary sequences.
  • normal alleles is meant the alleles as they have been isolated or as they can be isolated from samples of normal individuals, the “mutated alleles” being the alleles of the genes carrying the sequences [CAG] n or [CGG] n abnormally repeated.
  • sequences in question are for the first time identified as being part of a gene which may have a mutation, said genes not having, moreover, in themselves never been described.
  • the highlighted sequences show very variable percentages of heterozygosity (HTZ) ranging from 0 to 0.75%.
  • these sequences are identified in the table and can, if necessary, be isolated using the following primers: SEQ ED 1 to 24, primers 1 to 24 corresponding, in pairs, to sequences A to L mentioned above.
  • sequences thus highlighted can, first of all, be used within the framework of the diagnosis, and more exactly of a prognosis. Indeed, like a large number of diseases having a genetic support, the demonstration of the presence of a sequence presenting an abnormal number of repetitions of triplet [CAG] n or [CGG] n cannot in itself ensure the occurrence of the disease, but must be interpreted according to a set of other information to allow either a very early diagnosis or possibly specific surveillance, especially in families at risk.
  • This diagnosis can be made by comparing the DNA sequence according to the patient's invention with a normal sequence to detect the presence of additional trinucleotide repeats. More particularly, the present invention relates to a method of highlighting the risks of the appearance of a disease trinucleotide repeat in a patient, characterized in that the DNA sequence according to the invention of said patient is compared to a normal sequence to detect the presence of additional trinucleotide repeats.
  • primers which can be used in the context of the methods according to the invention, mention should be made of the primers of SEQ IDs 1 to 24 which constitute pairs of primers for each of the sequences which are the subject of the invention.
  • sequences according to the present invention are amplified and it is then possible, by comparison with a normal and / or standard sample, to demonstrate the presence of the supernumerary triplets and therefore the possibility of occurrence of a disease linked to this type of dynamic mutation.
  • the present invention also relates to genes and their alleles which carry, at least in part, these sequences, said genes being, of course, directly involved in the occurrence of the disease. It also relates to a transformed cell expressing all or part of the above genes, a protein obtained from such a cell, a protein interacting with the aforementioned protein as well as a vector expressing one of these proteins.
  • the corresponding genes can be expressed in cells by known means in order to produce the corresponding proteins.
  • these proteins being involved in the disease, we will wish to reduce their quantity, either by blocking their expression by appropriate methods at the level of genes or regulatory elements, or by fixing or inactivating them, for example by using receptor proteins acting as decoys. Again, these receptor or inactivated proteins can be generated in situ by expression of the corresponding DNA sequences.
  • the second library contains 40,128 normalized cDNAs of newborn brains (NIB clones) subcloned into a lafmid vector (6) which is part of the resources of the IMAGE consortium
  • [CAG] n or [CGG] n perfect, but also the complex [CAG] n or [CGG] n sequences, that is to say those which are punctuated by insertions of triplets; indeed, this presence of inserted triples is observed in particular in the case of SCA1, while such is not the case for SBMA, MD and HD.
  • the new clones selected from NIB libraries are distinct from those selected from the FB group. This is in keeping with the fact that the human brain expresses a large number of distinct genes at different stages of development. Finally, from the present observations, it appears that [CAG] n or [CGG] n sequences are rare in human cDNAs representative of the 3 ′ regions of the mRNAs.

Abstract

The invention concerns a transcribed DNA sequence rich in CAG/CTG or CGG/GCC triplet repeat corresponding to sequences A to L according to the table and their alleles and the complementary sequences. Said sequences are useful in particular in the diagnosis of trinucleotide repeat disorders.

Description

SEQUENCES D'ADN RICHES EN TRIPLET REPETE UTILES DANS LE DIAGNOSTIC DE MALADIES A REPETITION TRTNUCLEOTIDIQUE REPEATED TRIPLET-RICH DNA SEQUENCES USEFUL IN THE DIAGNOSIS OF TRTNUCLEOTIDE REPEATED DISEASES
La présente invention concerne la mise en évidence de gènes humains comportant des séquences à triplets répétés CAG/CTG ou CGG/GCC, ainsi que l'utilisation de ces gènes dans le diagnostic et l'éventuel traitement de certaines maladies monogéniques ou maladies polygéniques à composante héréditaire.The present invention relates to the detection of human genes comprising sequences with repeated triplets CAG / CTG or CGG / GCC, as well as the use of these genes in the diagnosis and the possible treatment of certain monogenic diseases or polygenic diseases with component hereditary.
Le polymorphisme anormal de séquences à triplets répétés CAG/CTG ou CGG/GCC constitue une mutation impliquée dans au moins dix maladies humaines héréditaires (appelées maladies à triplet répété) qui sont principalement, mais non exclusivement, des maladies neurodégénératives. L'expansion (plusieurs triplets surnuméraires chez les malades) de séquences à triplet répété CAG/CTG est impliquée notamment dans l'atrophie spinobulbaire (SBMA), la dystrophie myotonique (DM), l'ataxie spinocérébelleuse (SCA) SCA1, SCA2, SCA3, et SCA6, l'atrophie dendato-rubro-pallydoluysiane (DRPLA), et la maladie de Huntington (1 à 3). L'expansion de séquences à triplet répété CGG/GCC est notamment impliquée dans le syndrome du X fragile (1). Enfin, le polymorphisme restreint de séquences à triplet répété CAG/CTG (par exemple un seul triplet surnuméraire chez les malades) est notamment impliquée dans la maladie de Behçet, une maladie autoimmune pour laquelle le passage de 5 à 6 codons GCT codant pour une polyalanine est associé à la maladie (4).The abnormal polymorphism of CAG / CTG or CGG / GCC repeating triplet sequences is a mutation involved in at least ten inherited human diseases (called repeating triplet diseases) that are primarily, but not exclusively, neurodegenerative diseases. The expansion (several supernumerary triplets in patients) of repeated CAG / CTG triplet sequences is involved in particular in spinobulbar atrophy (SBMA), myotonic dystrophy (DM), spinocerebellar ataxia (SCA1) SCA1, SCA2, SCA3 , and SCA6, dendato-rubro-pallydoluysiane atrophy (DRPLA), and Huntington's disease (1 to 3). The expansion of CGG / GCC repeat triplet sequences is notably implicated in fragile X syndrome (1). Finally, the restricted polymorphism of sequences with a repeated CAG / CTG triplet (for example a single supernumerary triplet in patients) is notably implicated in Behçet's disease, an autoimmune disease for which the passage of 5 to 6 GCT codons coding for a polyalanine is associated with the disease (4).
L'expansion des CAG/CTG ou CGG/GCC (appelée également mutation dynamique) est souvent associée à un phénomène d'anticipation, la taille des répétitions étant souvent corrélée de façon inverse avec l'âge de la survenue et/ou la sévérité des symptômes.The expansion of CAG / CTG or CGG / GCC (also called dynamic mutation) is often associated with a phenomenon of anticipation, the size of the repetitions often being inversely correlated with the age of onset and / or the severity of symptoms.
L'expansion des répétitions CAG/CTG peut apparaître dans les régions non codantes (DM) ou codantes (par exemple SBMA, HD, DRPLA, SCAs) des transcrits. Ces transcrits sont parfois exprimés dans les tissus autres que le cerveau et lorsqu'ils sont traduits conduisent à des produits de gènes plus grands et portant un domaine polyglutamine anormalement allongé (1 à 3). L'expansion de triplets répétés CAG/CTG peut être très importante (par exemple DM ; plusieurs centaines de triplets), modérée (par exemple HD ou SCA1 ; en général plus de 35 triplets), voire très modérée (par exemple SCA6 ; en général moins de 35 triplets). A l'état normal, ces répétitions peuvent être hautement polymoφhiquesThe expansion of the CAG / CTG repeats can appear in the non-coding (DM) or coding regions (for example SBMA, HD, DRPLA, SCAs) of the transcripts. These transcripts are sometimes expressed in tissues other than the brain and when they are translated lead to products of larger genes and carrying an abnormally elongated polyglutamine domain (1 to 3). The expansion of repeated CAG / CTG triplets can be very large (for example DM; several hundred triplets), moderate (for example HD or SCA1; in general more than 35 triplets), or even very moderate (for example SCA6; in general less than 35 triples). In the normal state, these repetitions can be highly polymorphic
(par exemple HD ou SCA1) ou faiblement à très faiblement polymorphiques (par exemple SCA2 et SCA6) (1 à 3). Une variation d'un seul triplet à l'intérieur d'un triplet répété court et faiblement polymoφhique peut aussi causer une maladie (4).(for example HD or SCA1) or weakly to very weakly polymorphic (for example SCA2 and SCA6) (1 to 3). A variation of a single triplet within a short, weakly polymoφhical repeated triplet can also cause disease (4).
La mise en évidence d'expansion CAG/CTG dans PADN génomique de patients (5) et/ou l'anticipation dans les familles à risque a suggéré que les mutations dynamiques sont impliquées dans SCA4, SCA5, l'ataxie cérébrale dominante autosomale (ADCA de type 2, aussi désignée sous le sigle SCA7), et dans les forme familiales de la psychose maniaco-dépressive (PMD) et de la schizophrénie (6, 7). Dans le cas de SCA7, la mise en évidence d'expansions de polyglutamine a suggéré que le triplet répété impliqué est du type triplet répétéEvidence of CAG / CTG expansion in patients' genomic DNA (5) and / or anticipation in families at risk suggested that dynamic mutations are involved in SCA4, SCA5, autosomal dominant cerebral ataxia (ADCA) type 2, also known as SCA7), and in familial forms of manic-depressive psychosis (PMD) and schizophrenia (6, 7). In the case of SCA7, the evidence of polyglutamine expansions suggested that the repeat triplet involved is of the repeat triplet type.
CAG dans une région codante (8).CAG in a coding region (8).
L'autisme et la démence familiale qui montrent des variabilités dans l'âge de la survenue ou dans la sévérité des symptômes pourraient également être causées par des mutations dynamiques. Un assez grand nombre de maladies pourraient également avoir pour origine ou impliquer des mutations dynamiques : la maladie de Parkinson, les paraplégies spastiques, les ataxies cérébrospinale non répertoriées précédemment, - les cataractes zonulaires, les tremblements incontrôlables, les neuropathies amyloïdes familiales, les arthrites granulomateuses familiales (maladie de Blau), les microsomies hémifaciales, - certaines anémies et glaucomes, les désordres obsessionnels. •Ce qui précède démontre l'importance considérable des triplets répétés pour le diagnostic et le traitement de plusieurs maladies.Autism and familial dementia, which show variability in the age of onset or in the severity of symptoms, could also be caused by dynamic mutations. A fairly large number of diseases could also originate or involve dynamic mutations: Parkinson's disease, spastic paraplegias, cerebrospinal ataxias not previously listed, - zonular cataracts, uncontrollable tremors, familial amyloid neuropathies, granulomatous arthritis familial (Blau's disease), hemifacial microsomies, - certain anemias and glaucomas, obsessive-compulsive disorders. • The above demonstrates the considerable importance of repeated triplets for the diagnosis and treatment of several diseases.
La présente invention repose sur une étude systématique des répétitions CAG/CTG ou CGG/GCC (qui seront ci-après désignées par "[CAG]n" et "[CGG] n" respectivement, n étant le nombre de répétitions du triplet), cette étude ayant été réalisée avec des banques d'ADNc humains, celles-ci ayant subi un premier criblage général, puis les séquences retenues ayant été sélectionnées sur la base d'un certain nombre de critères permettant de s'assurer que les séquences en cause appartenaient à de nouveaux gènes humains et pouvaient, avec une grande probabilité, être impliquées dans des maladies à triplet répété. Enfin, les séquences sélectionnées ont fait l'objet d'une étude plus complète destinée à permettre leur localisation.The present invention is based on a systematic study of the CAG / CTG or CGG / GCC repeats (which will be designated below by "[CAG] n" and "[CGG] n" respectively, n being the number of repetitions of the triplet), this study having been carried out with human cDNA libraries, these having undergone a first general screening, then the selected sequences having been selected on the basis of a certain number of criteria making it possible to ensure that the sequences in question belonged to new human genes and could, with high probability, be implicated in repeated triplet diseases. Finally, the selected sequences were the subject of a more complete study intended to allow their localization.
Dans le cadre de la présente invention, deux ensembles d'ADNc provenant de cerveaux humains ont été étudiés pour analyser la présence de répétitions CAG en utilisant l'hybridation d'oligonucléotides sur des membranes à haute densité. Les deux librairies ont été obtenues à partir de ARNms à l'aide d'amorce oligo-dT, l'une des librairies étant constituée de clones d'ADNc de cerveau foetal (FB) et l'autre librairie étant constituée de clones d'ADNc normalisés de cerveau de nouveau-né (NIB). De façon générale, la présente invention repose sur la mise en évidence de certaines séquences susceptibles d'être impliquées dans les maladies à triplet répété obtenues dans des conditions d'hybridation permettant de sélectionner des ADNc qui contiennent un nombre de séquences [CAG] ou [CGG] supérieur à 7, lesquelles sont plus susceptibles d'être polymoφhes dans une population malade. Les conditions complètes d'analyse et de sélection de ces différentes séquences ne seront pas détaillées ci-après, seuls seront fournis les éléments permettant de les localiser et de les réisoler grâce, notamment, aux amorces PCR qui seront décrites ci-après.In the context of the present invention, two sets of cDNA from human brains were studied to analyze the presence of CAG repeats using the hybridization of oligonucleotides on high density membranes. The two libraries were obtained from mRNAs using the oligo-dT primer, one of the libraries being made up of fetal brain (FB) cDNA clones and the other library being made up of clones of Normalized newborn brain cDNA (NIB). In general, the present invention is based on the demonstration of certain sequences capable of being involved in repeated triplet diseases obtained under hybridization conditions making it possible to select cDNAs which contain a number of [CAG] or [ CGG] greater than 7, which are more likely to be polymorphous in a sick population. The complete conditions for analysis and selection of these different sequences will not be detailed below, only the elements making it possible to locate and re-isolate them will be provided, in particular, using the PCR primers which will be described below.
Plus particulièrement, la présente invention concerne une séquence d'ADN transcrit riche en triplet répété CAG/CTG ou CGG/GCC correspondant aux séquences A à L du tableau, ainsi que leurs alleles normaux et mutés et les séquences complémentaires.More particularly, the present invention relates to a transcribed DNA sequence rich in repetitive triplet CAG / CTG or CGG / GCC corresponding to sequences A to L of the table, as well as their normal and mutated alleles and the complementary sequences.
Par "alleles normaux" on entend désigner les alleles tels qu'ils ont été isolés ou tels qu'il peuvent être isolés de prélèvements d'individus normaux, les "alleles mutés" étant les alleles des gènes portant les séquences [CAG]n ou [CGG]n anormalement répétées.By "normal alleles" is meant the alleles as they have been isolated or as they can be isolated from samples of normal individuals, the "mutated alleles" being the alleles of the genes carrying the sequences [CAG] n or [CGG] n abnormally repeated.
Il est important de noter que, bien que certaines de ces séquences soient, totalement ou en partie, publiques, les séquences en cause sont pour la première fois identifiées comme faisant partie d'un gène pouvant présenter une mutation, lesdits gènes n'ayant, par ailleurs, en eux-mêmes jamais été décrits.It is important to note that, although some of these sequences are, in whole or in part, public, the sequences in question are for the first time identified as being part of a gene which may have a mutation, said genes not having, moreover, in themselves never been described.
Les séquences mises en évidence présentent des pourcentages d'hétérozygotie (HTZ) très variables allant de 0 à 0,75 %. Les séquences monomoφhiques (pour lesquelles HTZ = 0) sont au moins autant susceptibles d'être impliquées dans des maladies à triplet répété que les séquences polymoφhes. Bien entendu, comme cela est mentionné précédemment, ces séquences sont identifiées dans le tableau et pourront, si besoin est, être réisolées en utilisant les amorces suivantes : SEQ ED 1 à 24, les amorces 1 à 24 correspondant, par paire, aux séquences A à L mentionnées précédemment.The highlighted sequences show very variable percentages of heterozygosity (HTZ) ranging from 0 to 0.75%. The monomorphic sequences (for which HTZ = 0) are at least as likely to be involved in diseases with repeated triplets as the polymorphic sequences. Of course, as mentioned above, these sequences are identified in the table and can, if necessary, be isolated using the following primers: SEQ ED 1 to 24, primers 1 to 24 corresponding, in pairs, to sequences A to L mentioned above.
Les séquences ainsi mises en évidence peuvent, tout d'abord, être utilisées dans le cadre du diagnostic, et plus exactement d'un pronostic. En effet, comme un grand nombre de maladies ayant un support génétique, la mise en évidence de la présence d'une séquence présentant un nombre de répétitions anormal de triplet [CAG]n ou [CGG]n ne peut en elle-même assurer de la survenue de la maladie, mais doit être interprétée en fonction d'un ensemble d'autres informations pour permettre, soit un diagnostic très précoce, soit éventuellement une surveillance spécifique, surtout dans les familles à risque.The sequences thus highlighted can, first of all, be used within the framework of the diagnosis, and more exactly of a prognosis. Indeed, like a large number of diseases having a genetic support, the demonstration of the presence of a sequence presenting an abnormal number of repetitions of triplet [CAG] n or [CGG] n cannot in itself ensure the occurrence of the disease, but must be interpreted according to a set of other information to allow either a very early diagnosis or possibly specific surveillance, especially in families at risk.
Ce diagnostic peut être effectué en comparant la séquence d'ADN selon l'invention du patient avec une séquence normale pour détecter la présence de répétitions trinucléotidiques supplémentaires. Plus particulièrement, la présente invention concerne un procédé de mise en évidence des risques d'apparition d'une maladie à répétition trinucléotidique chez un patient, caractérisé en ce qu'on compare la séquence d'ADN conforme à l'invention dudit patient à une séquence normale pour détecter la présence de répétitions trinucléotidiques supplémentaires.This diagnosis can be made by comparing the DNA sequence according to the patient's invention with a normal sequence to detect the presence of additional trinucleotide repeats. More particularly, the present invention relates to a method of highlighting the risks of the appearance of a disease trinucleotide repeat in a patient, characterized in that the DNA sequence according to the invention of said patient is compared to a normal sequence to detect the presence of additional trinucleotide repeats.
Bien entendu, il existe un grand nombre de méthodes qui permettent la mise en évidence de triplets surnuméraires par rapport à des séquences normales, par exemple il est possible de mettre en évidence des différences de poids moléculaires en utilisant des gels et une méthode de type RFLP. Mais, les méthodes les plus efficaces consistent à pratiquer préalablement à la mise en évidence des différences une opération d'amplification, par toute méthode appropriée, notamment la méthode dite "PCR", même si d'autres méthodes sont utilisables.Of course, there are a large number of methods which allow the detection of supernumerary triplets compared to normal sequences, for example it is possible to highlight differences in molecular weights using gels and an RFLP type method. . However, the most effective methods consist in carrying out an amplification operation, before highlighting the differences, by any suitable method, in particular the so-called "PCR" method, even if other methods can be used.
Il n'est pas nécessaire ici de décrire en détail la méthode PCR, celle-ci permet d'amplifier de façon considérable une séquence spécifique comprise entre deux séquences appelées "amorces".It is not necessary here to describe in detail the PCR method, it makes it possible to considerably amplify a specific sequence comprised between two sequences called "primers".
Parmi les amorces utilisables dans le cadre des procédés selon l'invention, il faut citer les amorces des SEQ ID 1 à 24 qui constituent deux par deux des paires d'amorces pour chacune des séquences objet de l'invention.Among the primers which can be used in the context of the methods according to the invention, mention should be made of the primers of SEQ IDs 1 to 24 which constitute pairs of primers for each of the sequences which are the subject of the invention.
En utilisant les amorces décrites précédemment, on amplifie les séquences selon la présente invention et il est alors possible, par comparaison avec un échantillon normal et/ou étalon, de mettre en évidence la présence des triplets surnuméraires et donc la possibilité de survenue d'une maladie liée à ce type de mutation dynamique.Using the primers described above, the sequences according to the present invention are amplified and it is then possible, by comparison with a normal and / or standard sample, to demonstrate the presence of the supernumerary triplets and therefore the possibility of occurrence of a disease linked to this type of dynamic mutation.
Il est également intéressant de noter que certaines maladies à triplet répété sont connues pour être associées à des variations modérées à très faibles du nombre de triplets répétés, comme par exemple, SCA6 et la maladie de Behçet. Dans ces conditions, la méthode diagnostic peut permettre, non seulement un bon pronostic de la survenue de la maladie, mais également d'évaluer le moment où cette maladie surviendra et/ou son éventuelle sévérité.It is also interesting to note that certain repeated triplet diseases are known to be associated with moderate to very small variations in the number of repeated triplets, such as, for example, SCA6 and Behçet's disease. Under these conditions, the diagnostic method can allow not only a good prognosis for the onset of the disease, but also to assess the time when this disease will occur and / or its possible severity.
La présente invention concerne également les gènes et leurs alleles qui portent, au moins en parties, ces séquences, lesdits gènes étant, bien entendu, impliqués directement dans la survenue de la maladie. Elle a également pour objet une cellule transformée exprimant tout ou partie des susdits gènes, une protéine obtenue à partie d'une telle cellule, une protéine interagissant avec la protéine précédemment mentionnée ainsi qu'un vecteur exprimant l'une de ces protéines.The present invention also relates to genes and their alleles which carry, at least in part, these sequences, said genes being, of course, directly involved in the occurrence of the disease. It also relates to a transformed cell expressing all or part of the above genes, a protein obtained from such a cell, a protein interacting with the aforementioned protein as well as a vector expressing one of these proteins.
Les gènes correspondant pourront être exprimés dans des cellules par des moyens connus afin de produire les protéines correspondantes.The corresponding genes can be expressed in cells by known means in order to produce the corresponding proteins.
Il est possible d'envisager l'utilisation desdites protéines dans certains kits de diagnostic par exemple. De façon générale, ces protéines étant impliquées dans la maladie, on souhaitera en diminuer la quantité, soit en bloquant leur expression par des méthodes appropriées au niveau des gènes ou des éléments de régulation, soit en les fixant ou en les inactivant, par exemple en utilisant des protéines réceptrices jouant le rôle de leurres. Là encore, ces protéines réceptrices ou inactivées pourront être générées in situ par expression des séquences d'ADN correspondantes.It is possible to envisage the use of said proteins in certain diagnostic kits for example. In general, these proteins being involved in the disease, we will wish to reduce their quantity, either by blocking their expression by appropriate methods at the level of genes or regulatory elements, or by fixing or inactivating them, for example by using receptor proteins acting as decoys. Again, these receptor or inactivated proteins can be generated in situ by expression of the corresponding DNA sequences.
Il est également possible de prévoir la réalisation d'anticoφs monoclonaux correspondant à ces protéines afin d'envisager le blocage desdites protéines lorsque cela est souhaité, l'ensemble de ces opérations pouvant être réalisé directement in vivo par exemple, en utilisant des techniques de thérapie génique, en particulier en utilisant des vecteurs qui porteront les séquences d'expression des gènes.It is also possible to provide for the production of monoclonal anticoφs corresponding to these proteins in order to envisage blocking said proteins when desired, all of these operations can be carried out directly in vivo for example, using therapy techniques gene, in particular using vectors that will carry the gene expression sequences.
On a mis en évidence le fait que les protéines présentant des domaines polyglutamines anormalement étendus avaient des propriétés d'aggrégation anormales, tant entre elles, qu'avec d'autres protéines (11, 12). Les aggrégats ainsi créés sont probablement impliqués dans la genèse des maladies à triplet répété, c'est pourquoi il est également possible de prévoir une thérapie dans laquelle les agents thérapeutiques viendraient empêcher l'aggrégation, soit en bloquant la molécule comme décrit précédemment, soit en se fixant à la molécule pour empêcher le rapprochement avec d'autres protéines.It has been demonstrated that proteins with abnormally large polyglutamine domains have abnormal aggregation properties, both among themselves and with other proteins (11, 12). The aggregates thus created are probably involved in the genesis of diseases with repeated triplets, this is why it is also possible to provide a therapy in which the therapeutic agents would prevent aggregation, either by blocking the molecule as described above, or by attaching to the molecule to prevent rapprochement with other proteins.
On peut, dans le cadre de la thérapie, prévoir d'utiliser des variants complets ou délétés de ces protéines, normales ou non (quant au domaine [CAG]n ou [CGG]n). •Les séquences selon la présente invention peuvent être obtenues grâce aux informations figurant au tableau et aux références qui y sont données et en utilisant les séquences d'amorce qui sont décrites dans les identificateurs de séquence ci- joints. Les deux librairies utilisées sont : une première librairie (5) contenant 60 000 ADNc non normalisés de cerveau foetal humain (clones FB) (Laboratoire duIn the context of therapy, provision may be made to use complete or deleted variants of these proteins, normal or not (as regards the [CAG] n or [CGG] n domain). • The sequences according to the present invention can be obtained thanks to the information appearing in the table and to the references which are given therein and by using the primer sequences which are described in the attached sequence identifiers. The two libraries used are: a first library (5) containing 60,000 non-normalized human fetal brain cDNAs (FB clones) (Laboratoire du
Dr. Hans Lehrach, Max Plank Institute for Molecular Genetics, Berlin,Dr. Hans Lehrach, Max Plank Institute for Molecular Genetics, Berlin,
Allemagne) sous-clonés dans le vecteur p-SPORT-1 (Life Technologies, Inc., Gaithesburg, MA, USA), et la seconde librairie contient 40 128 ADNc normalisés de cerveau de nouveau-né (clones NIB) sous-clonés dans un vecteur lafmid (6) et qui est une partie des ressources du consortium IMAGEGermany) subcloned into the vector p-SPORT-1 (Life Technologies, Inc., Gaithesburg, MA, USA), and the second library contains 40,128 normalized cDNAs of newborn brains (NIB clones) subcloned into a lafmid vector (6) which is part of the resources of the IMAGE consortium
(Lawrence Livermore Lab., Livermore, CN) et du programme EST Merck WU (Washington University, St-Louis, LO, USA).(Lawrence Livermore Lab., Livermore, CN) and the EST Merck WU program (Washington University, St-Louis, LO, USA).
D'autre part, un certain nombre d'autres informations ont été recueillies en utilisant Genbank Database (NCBI Bethesda, MA, USA).On the other hand, a number of other information was collected using the Genbank Database (NCBI Bethesda, MA, USA).
Les méthodes de sélection et d'analyse qui ont été mises en oeuvre ne font pas en elles-mêmes partie de la présente invention puisque l'invention a essentiellement pour objet les séquences ainsi obtenues et leur utilisation, notamment, dans des méthodes de diagnostic ou des méthodes de traitement thérapeutique.The selection and analysis methods which have been implemented do not in themselves form part of the present invention since the invention essentially relates to the sequences thus obtained and their use, in particular, in diagnostic or methods of therapeutic treatment.
Dans la présente analyse, on a retenu, non seulement les séquencesIn this analysis, we have chosen not only the sequences
[CAG]n ou [CGG]n parfaites, mais également les séquences [CAG]n ou [CGG]n complexes, c'est-à-dire celles qui sont ponctuées par des insertions de triplets ; en effet cette présence de triplets insérés s'observe notamment dans le cas de SCA1, alors que tel n'est pas le cas pour SBMA, MD et HD.[CAG] n or [CGG] n perfect, but also the complex [CAG] n or [CGG] n sequences, that is to say those which are punctuated by insertions of triplets; indeed, this presence of inserted triples is observed in particular in the case of SCA1, while such is not the case for SBMA, MD and HD.
D'autre part, les nouveaux clones sélectionnés à partir de librairies NIB sont distincts de ceux sélectionnés dans le groupe FB. Ceci est en accord avec le fait que le cerveau humain exprime un grand nombre de gènes distincts à des stades de développement différents. Enfin, des présentes observations il ressort que des séquences [CAG]n ou [CGG]n sont rares dans les ADNc humains représentatifs des régions 3' des ARNm.On the other hand, the new clones selected from NIB libraries are distinct from those selected from the FB group. This is in keeping with the fact that the human brain expresses a large number of distinct genes at different stages of development. Finally, from the present observations, it appears that [CAG] n or [CGG] n sequences are rare in human cDNAs representative of the 3 ′ regions of the mRNAs.
Bien que dans l'état actuel de la présente invention, les séquences qui constituent l'objet de l'invention n'aient pas été reliées directement à des affections héréditaires, la présence d'extensions anormales CAG ou CGG peut être reliée avec le risque de survenue d'une maladie, en particulier lorsqu'il y a une composante familiale.Although in the present state of the present invention, the sequences which constitute the object of the invention have not been directly linked to hereditary disorders, the presence of abnormal CAG or CGG extensions may be linked with the risk illness, especially when there is a family component.
Il existe un assez grand nombre de maladies qui actuellement n'ont pas été localisées et trouveront probablement à être reliées avec les séquences selon la présente invention.There are quite a number of diseases which currently have not been localized and will likely find to be related to the sequences according to the present invention.
Parmi les éléments additionnels qui ont été pris en compte pour étudier la pertinence des séquences retenues figure l'existence éventuelle d'un cadre de lecture ouvert (ORF) dans lequel les séquences [CAG]n ou [CGG]n codent pour une extension polyglutamine. Among the additional elements which have been taken into account to study the relevance of the sequences selected is the possible existence of an open reading frame (ORF) in which the sequences [CAG] n or [CGG] n code for a polyglutamine extension .
TABLEAUBOARD
CDCD
* structure imparfaite * imperfect structure
LISTE DE SEQUENCESLIST OF SEQUENCES
(1) INFORMATIONS GENERALES:(1) GENERAL INFORMATION:
(î) DEPOSANT:(î) DEPOSITOR:
(A) NOM: FONDATION JEAN DAUSSET 6 CENTRE D'ETUDES DU(A) NAME: JEAN DAUSSET FOUNDATION 6 CENTER D'ETUDES DU
POLYMORPHISME HUMAIN (CEPH)HUMAN POLYMORPHISM (CEPH)
(B) RUE: 27 RUE JULIETTE DODU(B) STREET: 27 RUE JULIETTE DODU
(C) VILLE: PARIS(C) CITY: PARIS
(E) PAYS: FRANCE(E) COUNTRY: FRANCE
(F) CODE POSTAL: 75010(F) POSTAL CODE: 75010
(il) TITRE DE L' INVENTION: SEQUENCES D'ADN RICHES EN TRIPLET REPETE UTILES DANS LE DIAGNOSTIC DE MALADIES A REPETITION TRINUCLEOTIDIQUE(II) TITLE OF THE INVENTION: REPEATED TRIPLET-RICH DNA SEQUENCES USEFUL IN THE DIAGNOSIS OF TRINUCLEOTIDE DISEASE DISEASES
(in) NOMBRE DE SEQUENCES: 24(in) NUMBER OF SEQUENCES: 24
( v) FORME DECHIFFRABLE PAR ORDINATEUR:(v) COMPUTER-DETACHABLE FORM:
(A) TYPE DE SUPPORT: Floppy disk(A) TYPE OF SUPPORT: Floppy disk
(B) ORDINATEUR: IBM PC compatible(B) COMPUTER: IBM PC compatible
(C) SYSTEME D' EXPLOITATION: PC-DOS/MS-DOS(C) OPERATING SYSTEM: PC-DOS / MS-DOS
(D) LOGICIEL: Patentln Release {.1.0, Version #1.30 (OEB)(D) SOFTWARE: Patentln Release {.1.0, Version # 1.30 (EPO)
(2) INFORMATIONS POUR LA SEQ ID NO: 1:(2) INFORMATION FOR SEQ ID NO: 1:
(î) CARACTERISTIQUES DE LA SEQUENCE:(î) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 17 paires de bases(A) LENGTH: 17 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
( i) TYPE DE MOLECULE: ADNc(i) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lenrach, 3erlm, AllemagneLaboratory of Dr hans Lenrach, 3erlm, Germany
(B) CLONE: ICRFp507K15223(B) CLONE: ICRFp507K15223
(îx) CARACTERISTIQUE:(îx) CHARACTERISTIC:
(A) NOM/CLE: 1.45(A) NAME / KEY: 1.45
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 1: CGCTCCTTCC TCTCAAG 17(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 1: CGCTCCTTCC TCTCAAG 17
(2) INFORMATIONS POUR LA SEQ ID NO: 2:(2) INFORMATION FOR SEQ ID NO: 2:
(1) CARACTERISTIQUES DE LA SEQUENCE:(1) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 18 paires de bases(A) LENGTH: 18 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(il) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lenrach, Berlin, AllemagneLaboratory of Dr hans Lenrach, Berlin, Germany
(B) CLONE: ICRFp507K15223 (ix) CARACTERISTIQUE:(B) CLONE: ICRFp507K15223 (ix) CHARACTERISTIC:
(A) NOM/CLE: 1.45(A) NAME / KEY: 1.45
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 2: CTCAGCATGG GGTAAAAG 18(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 2: CTCAGCATGG GGTAAAAG 18
(2) INFORMATIONS POUR LA SEQ ID NO: 3:(2) INFORMATION FOR SEQ ID NO: 3:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 19 paires de bases(A) LENGTH: 19 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(ii) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vii) SOURCE IMMEDIATE:(vii) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lehrach, Berlin, AllemagneLaboratory of Dr hans Lehrach, Berlin, Germany
(B) CLONE: ICRFp507M22190(B) CLONE: ICRFp507M22190
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 2.52(A) NAME / KEY: 2.52
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 3: CTACAACAGA AATTCCGAC 19(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 3: CTACAACAGA AATTCCGAC 19
(2) INFORMATIONS POUR LA SEQ ID NO: 4:(2) INFORMATION FOR SEQ ID NO: 4:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 16 paires de bases(A) LENGTH: 16 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(ii) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vii) SOURCE IMMEDIATE:(vii) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lehrach, Berlin, AllemagneLaboratory of Dr hans Lehrach, Berlin, Germany
(B) CLONE: ICRFp507M22190(B) CLONE: ICRFp507M22190
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 2.52(A) NAME / KEY: 2.52
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 4: GGAACGACTG GAACAG 16(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 4: GGAACGACTG GAACAG 16
(2) INFORMATIONS POUR LA SEQ ID NO: 5:(2) INFORMATION FOR SEQ ID NO: 5:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 21 paires de bases(A) LENGTH: 21 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(ii) TYPE DE MOLECULE: ADNc (vu) SOURCE IMMEDIATE:(ii) TYPE OF MOLECULE: cDNA (seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lehrach, Berlin, AllemagneLaboratory of Dr hans Lehrach, Berlin, Germany
(B) CLONE: ICRF p507F12249(B) CLONE: ICRF p507F12249
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 2.99(A) NAME / KEY: 2.99
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 5: CATGGGGTTG CCGAAGAGGA G 21(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 5: CATGGGGTTG CCGAAGAGGA G 21
(2) INFORMATIONS POUR LA SEQ ID NO: 6:(2) INFORMATION FOR SEQ ID NO: 6:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 20 paires de bases(A) LENGTH: 20 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(n) TYPE DE MOLECULE: ADNc(n) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lehrach, Berlin, AllemagneLaboratory of Dr hans Lehrach, Berlin, Germany
(B) CLONE: ICRFp507F12249(B) CLONE: ICRFp507F12249
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 2.99(A) NAME / KEY: 2.99
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 6: CGCCCTTCTG GAAGGCTCAG 20(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 6: CGCCCTTCTG GAAGGCTCAG 20
(2) INFORMATIONS POUR LA SEQ ID NO: 7:(2) INFORMATION FOR SEQ ID NO: 7:
(l) CARACTERISTIQUES DE LA SEQUENCE:(l) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 18 paires de bases(A) LENGTH: 18 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(il) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lehrach, Berlin, AllemagneLaboratory of Dr hans Lehrach, Berlin, Germany
(B) CLONE: ICRFp507C15296(B) CLONE: ICRFp507C15296
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 2.122(A) NAME / KEY: 2.122
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 7:(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 7:
TGGCAGAATG GATTTGTG 18TGGCAGAATG GATTTGTG 18
(2) INFORMATIONS POUR LA SEQ ID NO: 8:(2) INFORMATION FOR SEQ ID NO: 8:
(i) CARACTERISTIQUES DE LA SEQUENCE. (A) LONGUEUR: 19 paires de bases (B) TYPE: nucléotide(i) CHARACTERISTICS OF THE SEQUENCE. (A) LENGTH: 19 base pairs (B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(11) TYPE DE MOLECULE: ADNc(11) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lehrach, Berlin, AllemagneLaboratory of Dr hans Lehrach, Berlin, Germany
(B) CLONE: ICRFp507C15296(B) CLONE: ICRFp507C15296
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 2.122(A) NAME / KEY: 2.122
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 8: CCATCTTGAA GAAGTACTT 19(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 8: CCATCTTGAA GAAGTACTT 19
(2) INFORMATIONS POUR LA SEQ ID NO: 9:(2) INFORMATION FOR SEQ ID NO: 9:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 19 paires de bases(A) LENGTH: 19 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
( l) TYPE DE MOLECULE: ADNc(l) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lehrach, Berlin, AllemagneLaboratory of Dr hans Lehrach, Berlin, Germany
(B) CLONE: ICRFp507L13177(B) CLONE: ICRFp507L13177
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 3.58(A) NAME / KEY: 3.58
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 9: TCACTCCTCC ATTGTCTGC 19(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 9: TCACTCCTCC ATTGTCTGC 19
(2) INFORMATIONS POUR LA SEQ ID NO: 10:(2) INFORMATION FOR SEQ ID NO: 10:
(i) CARACTERISTIQUES DE LA SEQUENCE.(i) CHARACTERISTICS OF THE SEQUENCE.
(A) LONGUEUR: 18 paires de bases(A) LENGTH: 18 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
( l) TYPE DE MOLECULE: ADNc(l) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE.(seen) IMMEDIATE SOURCE.
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lehrach, Berlin, AllemagneLaboratory of Dr hans Lehrach, Berlin, Germany
(B) CLONE: ICRFp507L13177(B) CLONE: ICRFp507L13177
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 3.58(A) NAME / KEY: 3.58
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 10: CTGTCCGAGG AGGTATCG 18(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 10: CTGTCCGAGG AGGTATCG 18
(2) INFORMATIONS POUR LA SEQ ID NO: 11:(2) INFORMATION FOR SEQ ID NO: 11:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 18 paires de bases(A) LENGTH: 18 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(il) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Reseau I.M.A.G.E., Lawrence Livermore National Laboratories, Livermore, CA, USA.(A) LIBRARY: I.M.A.G.E. Network, Lawrence Livermore National Laboratories, Livermore, CA, USA.
(B) CLONE: 35570(B) CLONE: 35570
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 1.10(A) NAME / KEY: 1.10
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 11: ACCTTGTCCA GTCCATTG 18(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 11: ACCTTGTCCA GTCCATTG 18
(2) INFORMATIONS POUR LA SEQ ID NO: 12:(2) INFORMATION FOR SEQ ID NO: 12:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 21 paires de bases(A) LENGTH: 21 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(il) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Réseau I.M.A.G.E., Lawrence Livermore National Laboratories, Livermore, CA, USA.(A) LIBRARY: I.M.A.G.E. Network, Lawrence Livermore National Laboratories, Livermore, CA, USA.
(B) CLONE: 35570(B) CLONE: 35570
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 1.10(A) NAME / KEY: 1.10
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 12: GTTGTCTTTA TAAAACACAG A 21(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 12: GTTGTCTTTA TAAAACACAG A 21
(2) INFORMATIONS POUR LA SEQ ID NO: 13:(2) INFORMATION FOR SEQ ID NO: 13:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 17 paires de bases(A) LENGTH: 17 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(il) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Reseau I.M.A.G.E., Lawrence Livermore National Laboratories, Livermore, CA, USA.(A) LIBRARY: I.M.A.G.E. Network, Lawrence Livermore National Laboratories, Livermore, CA, USA.
(B) CLONE: 43014 (ix) CARACTERISTIQUE:(B) CLONE: 43014 (ix) CHARACTERISTIC:
(A) NOM/CLE: 1.12(A) NAME / KEY: 1.12
(x ) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 13: CCCAAGTCCC ACAATAG 17(x) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 13: CCCAAGTCCC ACAATAG 17
(2) INFORMATIONS POUR LA SEQ ID NO: 14:(2) INFORMATION FOR SEQ ID NO: 14:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 16 paires de bases(A) LENGTH: 16 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(il) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Reseau I.M.A.G.E., Lawrence Livermore National Laboratories, Livermore, CA, USA.(A) LIBRARY: I.M.A.G.E. Network, Lawrence Livermore National Laboratories, Livermore, CA, USA.
(B) CLONE: 43014(B) CLONE: 43014
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 1.12(A) NAME / KEY: 1.12
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 14: GCGTCAGTGG CAGAAG 16(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 14: GCGTCAGTGG CAGAAG 16
(2) INFORMATIONS POUR LA SEQ ID NO: 15:(2) INFORMATION FOR SEQ ID NO: 15:
(l) CARACTERISTIQUES DE LA SEQUENCE:(l) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 18 paires de bases(A) LENGTH: 18 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(il) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Reseau I.M.A.G.E., Lawrence Livermore National Laboratories, Livermore, CA, USA.(A) LIBRARY: I.M.A.G.E. Network, Lawrence Livermore National Laboratories, Livermore, CA, USA.
(B) CLONE: 54662(B) CLONE: 54662
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 1.34(A) NAME / KEY: 1.34
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 15: CCTTTATGAT CCTGGTTC 18(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 15: CCTTTATGAT CCTGGTTC 18
(2) INFORMATIONS POUR LA SEQ ID NO: 16:(2) INFORMATION FOR SEQ ID NO: 16:
( ) CARACTERISTIQUES DE LA SEQUENCE:() CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 18 paires de bases(A) LENGTH: 18 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(il) TYPE DE MOLECULE: ADNc (vii) SOURCE IMMEDIATE:(ii) TYPE OF MOLECULE: cDNA (vii) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Reseau I.M.A.G.E., Lawrence Livermore National Laboratories, Livermore, CA, USA.(A) LIBRARY: I.M.A.G.E. Network, Lawrence Livermore National Laboratories, Livermore, CA, USA.
(B) CLONE: 54662(B) CLONE: 54662
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: i.34(A) NAME / KEY: i.34
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 16: TGTGTTAGGT CCTAGAAG 18(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 16: TGTGTTAGGT CCTAGAAG 18
(2) INFORMATIONS POUR LA SEQ ID NO: 17:(2) INFORMATION FOR SEQ ID NO: 17:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 18 paires de bases(A) LENGTH: 18 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(ii) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vii) SOURCE IMMEDIATE:(vii) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Reseau I.M.A.G.E., Lawrence Livermore National Laboratories, Livermore, CA, USA.(A) LIBRARY: I.M.A.G.E. Network, Lawrence Livermore National Laboratories, Livermore, CA, USA.
(B) CLONE: 42315(B) CLONE: 42315
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: i.lll(A) NAME / KEY: i.lll
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 17: CCTCCAGCAG GGTATGGC 18(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 17: CCTCCAGCAG GGTATGGC 18
(2) INFORMATIONS POUR LA SEQ ID NO: 18:(2) INFORMATION FOR SEQ ID NO: 18:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 19 paires de bases(A) LENGTH: 19 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(ii) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vii) SOURCE IMMEDIATE:(vii) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Reseau I.M.A.G.E., Lawrence Livermore National Laboratories, Livermore, CA, USA.(A) LIBRARY: I.M.A.G.E. Network, Lawrence Livermore National Laboratories, Livermore, CA, USA.
(B) CLONE: 42315(B) CLONE: 42315
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: i.lll(A) NAME / KEY: i.lll
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 18: CGAACGGACC TGGGAACTG 19(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 18: CGAACGGACC TGGGAACTG 19
(2) INFORMATIONS POUR LA SEQ ID NO: 19:(2) INFORMATION FOR SEQ ID NO: 19:
(i) CARACTERISTIQUES DE LA SEQUENCE: (A) LONGUEUR: 18 paires de bases (B) TYPE: nucléotide(i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(11) TYPE DE MOLECULE: ADNc(11) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Reseau I.M.A.G.E., Lawrence Livermore National Laboratories, Livermore, CA, USA.(A) LIBRARY: I.M.A.G.E. Network, Lawrence Livermore National Laboratories, Livermore, CA, USA.
(B) CLONE: 52900(B) CLONE: 52900
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 1.129(A) NAME / KEY: 1.129
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 19: TCTGCCCCTG GAGGTGGG 18(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 19: TCTGCCCCTG GAGGTGGG 18
(2) INFORMATIONS POUR LA SEQ ID NO: 20:(2) INFORMATION FOR SEQ ID NO: 20:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 27 paires de bases(A) LENGTH: 27 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(n) TYPE DE MOLECULE: ADNc(n) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Reseau I.M.A.G.E., Lawrence Livermore National Laboratories, Livermore, CA, USA.(A) LIBRARY: I.M.A.G.E. Network, Lawrence Livermore National Laboratories, Livermore, CA, USA.
(B) CLONE: 52900(B) CLONE: 52900
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE. 1.129(A) NAME / KEY. 1.129
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 20: CTTCACTGGC GCTTTTTCTT CAGCTTC 27(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 20: CTTCACTGGC GCTTTTTCTT CAGCTTC 27
(2) INFORMATIONS POUR LA SEQ ID NO: 21:(2) INFORMATION FOR SEQ ID NO: 21:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 19 paires de bases(A) LENGTH: 19 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(n) TYPE DE MOLECULE: ADNc(n) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lehrach, Berlin, AllemagneLaboratory of Dr hans Lehrach, Berlin, Germany
(B) CLONE: ICRFp507P04188(B) CLONE: ICRFp507P04188
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 1.85(A) NAME / KEY: 1.85
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 21: GAACAGCCTA TCTCATTCC 19(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 21: GAACAGCCTA TCTCATTCC 19
(2) INFORMATIONS POUR LA SEQ ID NO: 22:(2) INFORMATION FOR SEQ ID NO: 22:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 32 paires de bases(A) LENGTH: 32 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(il) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lehrach, Berlin, AllemagneLaboratory of Dr hans Lehrach, Berlin, Germany
(B) CLONE: ICRFp507P041188(B) CLONE: ICRFp507P041188
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 1.85(A) NAME / KEY: 1.85
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 22: TCAGAAGTAA CTTGAATAAA TAATCATATT CG 32(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 22: TCAGAAGTAA CTTGAATAAA TAATCATATT CG 32
(2) INFORMATIONS POUR LA SEQ ID NO: 23:(2) INFORMATION FOR SEQ ID NO: 23:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 16 paires de bases(A) LENGTH: 16 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(il) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lenrach, Berlin, AllemagneLaboratory of Dr hans Lenrach, Berlin, Germany
(B) CLONE: ICRFp507B16262(B) CLONE: ICRFp507B16262
(ix) CARACTERISTIQUE:(ix) CHARACTERISTIC:
(A) NOM/CLE: 2.296(A) NAME / KEY: 2.296
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 23:(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 23:
CACTTGGAAT CCACAC 16CACTTGGAAT CCACAC 16
(2) INFORMATIONS POUR LA SEQ ID NO: 24:(2) INFORMATION FOR SEQ ID NO: 24:
(i) CARACTERISTIQUES DE LA SEQUENCE:(i) CHARACTERISTICS OF THE SEQUENCE:
(A) LONGUEUR: 16 paires de bases(A) LENGTH: 16 base pairs
(B) TYPE: nucléotide(B) TYPE: nucleotide
(C) NOMBRE DE BRINS: simple(C) NUMBER OF STRANDS: single
(D) CONFIGURATION: linéaire(D) CONFIGURATION: linear
(il) TYPE DE MOLECULE: ADNc(ii) TYPE OF MOLECULE: cDNA
(vu) SOURCE IMMEDIATE:(seen) IMMEDIATE SOURCE:
(A) BIBLIOTHEQUE: Max Plank Institute for Molecular Genetics,(A) LIBRARY: Max Plank Institute for Molecular Genetics,
Laboratory of Dr hans Lehrach, Berlin, AllemagneLaboratory of Dr hans Lehrach, Berlin, Germany
(B) CLONE: ICRFp507B16262 (ix) CARACTERISTIQUE:(B) CLONE: ICRFp507B16262 (ix) CHARACTERISTIC:
(A) NOM/CLE: 2.296(A) NAME / KEY: 2.296
(xi) DESCRIPTION DE LA SEQUENCE: SEQ ID NO: 24: AGAGATCCTG AAGGAC 16 (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 24: AGAGATCCTG AAGGAC 16
REFERENCESREFERENCES
1. Paulson, H.L. and Fishbeck, K.H. Trinucleotide repeats in neurogenetic disorders Annu Rev NeuroSci 19, 79-107 (1996). 2. Zohgbi, H. The expanding world of ataxins. Nature Genêt. 14, 237-1. Paulson, H.L. and Fishbeck, K.H. Trinucleotide repeats in neurogenetic disorders Annu Rev NeuroSci 19, 79-107 (1996). 2. Zohgbi, H. The expanding world of ataxins. Nature Broom. 14, 237-
238 (1996).238 (1996).
3. Zuchenko O, Bailey J, Bonnen P, Ashizawa T, Stockton DW, Amos C, Dobyns WB, Subramony SH, Zoghbi HY, and Lee CC. Autosomal dominant cerebellar ataxia (SCA6) associated with small glutamine expansion in the alpha lA-voltage-dependent calcium channel. Nature3. Zuchenko O, Bailey J, Bonnen P, Ashizawa T, Stockton DW, Amos C, Dobyns WB, Subramony SH, Zoghbi HY, and Lee CC. Autosomal dominant cerebellar ataxia (SCA6) associated with small glutamine expansion in the alpha lA-voltage-dependent calcium channel. Nature
Genêt 15, 62-69 (1997).Genet 15, 62-69 (1997).
4. Mizu i, N. et al. Triplet repeat polymorphism in the transmembrane région of the MICA gène: a strong association of six GCT répétitions with Behçet disease. Proc Natl Acad Sci USA 94, 1298-1303 (1997). 5. Schalling M, Hudson TJ, Buetow KH, Housman DE. Direct détection of novel expanded trinucleotide repeats in the human génome. Nature Genêt 4, 135-139, 1993.4. Mizu i, N. et al. Triplet repeat polymorphism in the transmembrane region of the MICA gene: a strong association of six GCT repeats with Behçet disease. Proc Natl Acad Sci USA 94, 1298-1303 (1997). 5. Schalling M, Hudson TJ, Buetow KH, Housman DE. Direct detection of novel expanded trinucleotide repeats in the human genome. Nature Broom 4, 135-139, 1993.
6. O'Donovan MC, Guy C, Craddock N, Murphy KC, Cardno AG, Jones LA, Oven MJ McGuffin P. Expanded CAG repeats in schizophrema and bipolar disorders. Nature Genêt 10, 380-381 (1995).6. O'Donovan MC, Guy C, Craddock N, Murphy KC, Cardno AG, Jones LA, Oven MJ McGuffin P. Expanded CAG repeats in schizophrema and bipolar disorders. Nature Broom 10, 380-381 (1995).
7. Morris AG, Gaitonde E, McKenna PJ, Mollon JD, Hunt DM. CAG repeat expansions and schizophrenia : association with disease in females and with early age-at-onset. Hum Mol Genêt 4, 1957-1961 (1995).7. Morris AG, Gaitonde E, McKenna PJ, Mollon JD, Hunt DM. CAG repeat expansions and schizophrenia: association with disease in females and with early age-at-onset. Hum Mol Genêt 4, 1957-1961 (1995).
8. Trottier, Y. et al. Polyglutamine expansion as a pathological epitope in Huntington's disease and four dominant cerebellar ataxia. Nature 378,8. Trottier, Y. et al. Polyglutamine expansion as a pathological epitope in Huntington's disease and four dominant cerebellar ataxia. Nature 378,
403-406 (1995).403-406 (1995).
9. Meier-Ewert, S., Maier, E., Ahmadi, A., Curtis, J. and Lebrach, H. An Automated approach to generating expressed séquence catalogues. Nature 361, 375 (1993). 10. Soares, M.B., Bonaldo, M.F., Jelene, P., Su, L., Lawton, L. & Efstratiadis, A. Construction and characterization of a normalized cDNA library. Proc. Natl. Acad. Sci. USA. 91, 9228-9232 (1994).9. Meier-Ewert, S., Maier, E., Ahmadi, A., Curtis, J. and Lebrach, H. An Automated approach to generating expressed sequence catalogs. Nature 361, 375 (1993). 10. Soares, MB, Bonaldo, MF, Jelene, P., Su, L., Lawton, L. & Efstratiadis, A. Construction and characterization of a normalized cDNA library. Proc. Natl. Acad. Sci. USA. 91, 9228-9232 (1994).
11. Haad, D., Perry, M.J. and Haynes, L. Cellular transglutaminases in neural development. Int. J. Devl Neuroscience 11, 709-720 (1993).11. Haad, D., Perry, M.J. and Haynes, L. Cellular transglutaminases in neural development. Int. J. Devl Neuroscience 11, 709-720 (1993).
12. Li X-J, Li S-H, Sharp AH, Nucifora Jr FC, Schilling G, Lanahan A, Woriey P, Snyder SH, Ross CA. A Hungtintin-associated protein enriched in brain with implications for pathology. Nature 378, 398-402 (1995). 12. Li X-J, Li S-H, Sharp AH, Nucifora Jr FC, Schilling G, Lanahan A, Woriey P, Snyder SH, Ross CA. A Hungtintin-associated protein enriched in brain with implications for pathology. Nature 378, 398-402 (1995).

Claims

REVENDICATIONS
1) Séquence d'ADN transcrit riche en triplet répété CAG/CTG ou CGG/GCC correspondant aux séquences A à L selon le tableau ainsi que leurs alleles normaux et mutés et les séquences complémentaires.1) Sequence of transcribed DNA rich in repeated triplet CAG / CTG or CGG / GCC corresponding to sequences A to L according to the table as well as their normal and mutated alleles and the complementary sequences.
2) Séquence d'ADN, caractérisée en ce qu'elle porte au moins en partie une séquence selon la revendication 1.2) DNA sequence, characterized in that it carries at least partially a sequence according to claim 1.
3) Procédé de mise en évidence des risques d'apparition d'une maladie à répétition trinucléotidique chez un patient, caractérisé en ce qu'on compare la séquence d'ADN selon la revendication 1 dudit patient à une séquence normale pour détecter la présence de répétitions trinucléotidiques supplémentaires.3) Method for highlighting the risks of the appearance of a trinucleotide recurrent disease in a patient, characterized in that the DNA sequence according to claim 1 of said patient is compared with a normal sequence to detect the presence of additional trinucleotide repeats.
4) Procédé selon la revendication 3, caractérisé en ce qu'on effectue une amplification de tout ou partie des séquences selon la revendication 1 et que l'on met en évidence dans le produit d'amplification la présence de répétitions trinucléotidiques supplémentaires.4) Method according to claim 3, characterized in that one carries out an amplification of all or part of the sequences according to claim 1 and that one highlights in the amplification product the presence of additional trinucleotide repeats.
5) Procédé selon la revendication 4, caractérisé en ce que l'amplification des séquences est effectuée avec les amorces décrites SEQ ID 1 à 24 correspondant aux séquences A à L respectivement.5) Method according to claim 4, characterized in that the amplification of the sequences is carried out with the primers described SEQ ID 1 to 24 corresponding to the sequences A to L respectively.
6) Cellule transformée exprimant tout ou partie d'un gène selon la revendication 2.6) A transformed cell expressing all or part of a gene according to claim 2.
7) Protéine obtenue à partir d'une cellule transformée selon la revendication 6.7) Protein obtained from a transformed cell according to claim 6.
8) Protéine interagissant avec une protéine selon la revendication 7.8) Protein interacting with a protein according to claim 7.
9) Anticorps monoclonal correspondant à une protéine selon l'une des revendications 7 ou 8.9) Monoclonal antibody corresponding to a protein according to one of claims 7 or 8.
10) Vecteur exprimant une protéine selon l'une des revendications 7 ou 8.10) Vector expressing a protein according to one of claims 7 or 8.
11) Utilisation d'une protéine selon l'une des revendications 7 ou 8, d'un anticorps selon la revendication 9 ou d'un vecteur selon la revendication 10 pour la préparation d'un médicament. 11) Use of a protein according to one of claims 7 or 8, an antibody according to claim 9 or a vector according to claim 10 for the preparation of a medicament.
EP98929536A 1997-06-11 1998-06-10 Dna sequences rich in triplet repeat useful in the diagnosis of trinucleotide repeat disorders Withdrawn EP0988399A1 (en)

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CA2252727A1 (en) * 1998-11-03 2000-05-03 Ridha Joober Polyglutamine containing proteins and use thereof for diagnosis and treatment of cag repeat-linked neuropsychiatric disorders
WO2001018544A1 (en) * 1999-09-09 2001-03-15 Mcgill University Diagnosis, prognosis and treatment of trinucleotide repeat-associated diseases and intranuclear inclusions-associated diseases
KR20030013019A (en) * 2001-08-06 2003-02-14 주식회사 바이오메드랩 Diagnosis kit and diagnosis apparatus for multiplication disease of trinucleotide repeated sequence
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GB9311113D0 (en) * 1993-05-28 1993-07-14 Medical Res Council Probes
US5834183A (en) * 1993-06-29 1998-11-10 Regents Of The University Of Minnesota Gene sequence for spinocerebellar ataxia type 1 and method for diagnosis
WO1995025179A1 (en) * 1994-03-17 1995-09-21 University Of Massachusetts Medical Center Detection of trinucleotide repeats by in situ hybridization
FR2745007B1 (en) * 1996-02-15 1998-05-07 Fondation Jean Dausset Ceph DIAGNOSIS OF TRINUCLEOTIDE REPEATED DISEASES AND GENES INVOLVED IN SUCH DISEASES
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