在BLM缺陷胚胎干细胞中使用遗传选确定不匹配修复基因
Ge Guo1, Wei Wang, Allan Bradley
1The Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK.
Nature
|June 25, 2004
概括
研究人员使用Bloom开发了一种新的方法,用于在培养细胞中产生同卵性突变,使用Bloom.
科学领域:
- 遗传学 是一个遗传学.
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 双胞胎生物中的递归遗传选需要同卵性突变.
- 在培养细胞中产生同卵性突变是具有挑战性的,限制了递归选.
- 胚胎干细胞 (ES) 提供了对哺乳动物细胞基因进行研究的机会.
研究的目的:
- 建立一种可靠的方法,在培养ES细胞中产生同卵性突变,用于衰退性遗传查.
- 开发一个全基因组的同卵性突变ES细胞库.
- 在DNA不匹配修复 (MMR) 中选此库的缺陷.
主要方法:
- 在布鲁姆综合征蛋白 (Blm) 缺乏ES细胞中利用了高的线粒重组率.
- 使用可逆基因陷逆转病毒,从异构基因突变中生成同卵性突变细胞.
- 在DNA不匹配修复 (MMR) 中检查突变细胞库的缺陷.
主要成果:
- 成功生成了一个全基因组的同卵性突变ES细胞库.
- 在已知和新型MMR基因中确定了同卵性突变.
- 鉴定Dnmt1为一种新的MMR基因,Dnmt1缺乏的细胞显示微卫星不稳定性.
结论:
- 缺乏Blm的ES细胞系统能够基于表型进行衰退性遗传选.
- 这种方法为研究哺乳动物细胞中的基因功能提供了一个新的工具.
- 这些发现阐明了Dnmt1在DNA不匹配修复和癌症中的作用.
相关概念视频
Mismatch Repair
Overview
In-vitro Mutagenesis
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
Long-patch Base Excision Repair
Since the discovery of the two BER pathways, there has been a debate about how a cell chooses one pathway over the other and the factors determining this selection. Numerous in vitro experiments have pointed out multiple determinants for the sub-pathway selection. These are:
Mismatch Repair
Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...


