A model system to study genomic imprinting of human genes
A model system to study genomic imprinting of human genes
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DOI：
10.1073/pnas.95.25.14857
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发表时间：
1998-12-08
影响因子：
11.1
通讯作者：
Nicholls, RD
中科院分区：
文献类型：
--
作者：
Gabriel, JM;Higgins, MJ;Nicholls, RD
体细胞杂交已被证明保持正确的表观遗传染色质状态，以研究发育珠蛋白基因表达以及活性和非活性X染色体上的基因表达。这表明，潜在的使用体细胞杂交包含一个母亲或父亲的人类染色体作为一个模型系统，研究已知的印迹基因，并确定尚未未知的印迹基因。通过使用逆转录PCR检测基因表达，我们发现，功能印记保持在四个先前表征的15 q11-q13基因座的杂交包含一个单一的人类15号染色体和两个染色体11 p15基因座的杂交包含一个单一的染色体II。相比之下，15 q12-q13中的三个γ-氨基丁酸A型受体亚基基因是非印迹的。此外，我们发现SNRPN启动子和11 p15中CpG岛的差异DNA甲基化印记也在体细胞杂交中保持。因此，体细胞杂交是一个有效的和强大的系统，用于研究已知的印记基因，以及快速识别新的印记基因。
Somatic-cell hybrids have been shown to maintain the correct epigenetic chromatin states to study developmental globin gene expression as well as gene expression on the active and inactive X chromosomes. This suggests the potential use of somatic-cell hybrids containing either a maternal or a paternal human chromosome as a model system to study known imprinted genes and to identify as-yet-unknown imprinted genes. Testing gene expression by using reverse transcription followed by PCR, we show that functional imprints are maintained at four previously characterized 15q11-q13 loci in hybrids containing a single human chromosome 15 and at two chromosome 11p15 loci in hybrids containing a single chromosome II. In contrast, three gamma-aminobutyric acid type A receptor subunit genes in 15q12-q13 are nonimprinted. Furthermore, we have found that differential DNA methylation imprints at the SNRPN promoter and at a CpG island in 11p15 are also maintained in somatic-cell hybrids. Somatic-cell hybrids therefore are a valid and powerful system for studying known imprinted genes as well as for rapidly identifying new imprinted genes.