原位邻近连接检测法用于监测细胞内的整体 DNA 甲基化。
Proximity ligation in situ assay for monitoring the global DNA methylation in cells.
机构信息
Centre de Recherche en Cancérologie Nantes-Angers, INSERM, U892, Equipe Apoptose et Progression Tumorale, Equipe labellisée Ligue Nationale Contre le Cancer, 8 quai moncousu, BP7021, 44007 Nantes, France.
出版信息
BMC Biotechnol. 2011 Apr 6;11:31. doi: 10.1186/1472-6750-11-31.
BACKGROUND
DNA methylation has a central role in the epigenetic control of mammalian gene expression, and is required for X inactivation, genomics imprinting and silencing of retrotransposons and repetitive sequences. Thus, several technologies have been developed to measure the degree of DNA methylation.
RESULTS
We here present the development of the detection of protein-protein interactions via the adaptation of the proximity ligation in situ technology to evaluate the DNA methylation status in cells since the quantification of Dnmt1/PCNA interaction in cells reflects the degree of DNA methylation.
CONCLUSION
This method being directly realizable on cells, it appears that it could suggest a wide range of applications in basic research and drug development. More particularly, this method is specially adapted for the investigations realized from a weak quantity of biologic materiel such as stem cells or primary cultured tumor cells for examples.
背景
DNA 甲基化在哺乳动物基因表达的表观遗传调控中起着核心作用，对于 X 染色体失活、基因组印迹以及逆转录转座子和重复序列的沉默都是必需的。因此，已经开发了几种技术来测量 DNA 甲基化的程度。
结果
我们在这里介绍了通过适应原位邻近连接技术来检测蛋白质-蛋白质相互作用的方法，以评估细胞中的 DNA 甲基化状态，因为细胞中 Dnmt1/PCNA 相互作用的定量反映了 DNA 甲基化的程度。
结论
由于该方法可以直接在细胞上实现，因此它似乎可以在基础研究和药物开发等领域中得到广泛应用。特别是，该方法特别适用于从少量生物材料（例如干细胞或原代培养的肿瘤细胞）进行的研究。
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