Genome editing. The mutagenic chain reaction: a method for converting heterozygous to homozygous mutations.
Genome editing. The mutagenic chain reaction: a method for converting heterozygous to homozygous mutations.
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DOI：
10.1126/science.aaa5945
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发表时间：
2015-04-24
期刊：
影响因子：
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通讯作者：
Bier E
中科院分区：
文献类型：
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作者：
Gantz VM;Bier E
具有单个隐性功能丧失等位基因的生物体通常将具有野生型表型，而对于该等位基因的两个拷贝纯合的个体将显示突变体表型。在这里，我们开发了一种称为诱变链式反应（MCR）的方法，该方法基于CRISPR/Cas9基因组编辑系统，用于产生自催化突变以产生纯合功能丧失突变。我们在果蝇中证明，MCR突变有效地从其起源染色体传播到同源染色体，从而在绝大多数体细胞和生殖细胞中将杂合突变转化为纯合突变。MCR技术应该在不同的生物体中有广泛的应用。
An organism with a single recessive loss-of-function allele will typically have a wild-type phenotype while individuals homozygous for two copies of the allele will display a mutant phenotype. Here, we develop a method that we refer to as the mutagenic chain reaction (MCR), which is based on the CRISPR/Cas9 genome editing system for generating autocatalytic mutations to generate homozygous loss-of-function mutations. We demonstrate in Drosophila that MCR mutations efficiently spread from their chromosome of origin to the homologous chromosome thereby converting heterozygous mutations to homozygosity in the vast majority of somatic and germline cells. MCR technology should have broad applications in diverse organisms.