清脆的
Cas9
生物
基因组编辑
效应器
质粒
基因
计算生物学
DNA
引导RNA
基因靶向
基因组
遗传学
基因敲除
CRISPR干扰
细胞生物学
作者
Eric Ehrke-Schulz,Maren Schiwon,Claudia Hagedorn,Anja Ehrhardt
出处
期刊:Methods in molecular biology
日期:2017-01-01
被引量:3
标识
DOI:10.1007/978-1-4939-7231-9_11
摘要
CRISPR/Cas9 RNA-guided nucleases refashioned in vivo gene editing approaches for specific gene disruption, gene correction, or gene addition. Moreover, chimeric Cas9 proteins can be applied to direct fused cis-acting effector protein domains, enzymes, or fluorescent markers to DNA to target sequences to regulate gene expression, to introduce epigenetic changes, or to fluorescently label DNA sequences of interest. Here we show how to design guide RNAs for specific DNA targeting. We provide a protocol to customize the CRISPR/Cas9 machinery encoded on commercially available plasmids and present how to test the targeting efficiency of Cas9 with a target-specific gRNA by testing mutation induction efficiency. To exemplify related applications we provide a guideline of how to apply the CRISPR/Cas9 technology for gene labeling.
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