在小鼠肝脏中,CRISPR介导癌症基因的直接突变
Wen Xue1, Sidi Chen1, Hao Yin1
11] David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02142, USA [2].
Nature
|August 15, 2014
概括
研究人员开发了一种用于小鼠体内基因编辑的新型CRISPR/Cas系统,通过直接突变Pten和p53.3等瘤抑制基因,使得肝癌模型的快速生成成为可能.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 癌症研究 癌症研究
背景情况:
- 传统的小鼠癌症模型依赖于转基因或胚胎干细胞基因向.
- 开发用于体内癌症建模的新方法对于功能基因组学至关重要.
研究的目的:
- 建立一种新的CRISPR/Cas系统,用于在野生类型小鼠中体内生成肝癌模型.
- 为了证明在肝脏中直接突变瘤抑制基因和瘤基因的可行性.
主要方法:
- 在体内利用水力动力注射提供CRISPR/Cas9等离子体和单向导RNA (sgRNAs),准Pten和p53.
- 与sgRNA和DNA捐赠者寡核酸一起注入Cas9等离子体,在β-catenin基因中引入特定突变.
主要成果:
- 通过CRISPR介导的Pten突变产生了复制基因删除效应,导致脂质积累和Akt酸化升高.
- 同时Pten和p53向诱导的肝脏瘤,类似于Cre-LoxP模型中的瘤.
- 在生成的肝脏瘤中确认了Pten和p53的双基突变.
- 通过向基因编辑成功生成具有核β-catenin定位的肝细胞.
结论:
- 克里斯普尔/卡斯系统为肝脏癌症基因的直接突变提供了一种可行且快速的方法.
- 这种方法为快速开发肝癌模型和功能基因组学研究提供了新的途径.
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