Bisulfite genomic sequencing of microdissected cells.
Mapping of methylation patterns in CpG islands has become an important tool for understanding tissue-specific gene expression in both normal and pathological situations. However, the inherent cellular heterogeneity of any given tissues can affect the outcome and interpretation of molecular studies. In order to analyse genomic DNA methylation on a pure cell population from tissue sample, we have developed a simple technique of single-cell microdissection from cryostat sections which can be combined with bisulfite-mediated sequencing of 5-methylcytosine. We report here our results on the methylation status of the androgen receptor gene studied by bisulfite genomic sequencing on purified cells isolated from human testis.
Kerjean
A
A
Institut Cochin de Génétique Moléculaire, Laboratoire de Biologie de la Reproduction, Hôpital Cochin, 75014 Paris, France. kerjean@cochin.inserm.fr
Vieillefond
A
A
Thiounn
N
N
Sibony
M
M
Jeanpierre
M
M
Jouannet
P
P
D002478:Cells, Cultured; D018899:CpG Islands / Q000235:genetics; D004247:DNA / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D004210:Dissection / Q000379:methods; D005091:Exons / Q000235:genetics; D016678:Genome; D023281:Genomics / Q000379:methods; D006801:Humans; D008297:Male; D011110:Polymorphism, Genetic / Q000235:genetics; D011944:Receptors, Androgen / Q000235:genetics*; D012685:Sepharose; D017422:Sequence Analysis, DNA / Q000379:methods*; D013447:Sulfites / Q000378:metabolism*; D013737:Testis / Q000166:cytology / Q000378:metabolism / Q000473:pathology
D011944:Receptors, Androgen; D013447:Sulfites; D004247:DNA; D012685:Sepharose