Generating Custom Pooled CRISPR Libraries for Genetic Dissection of Biological Pathways.
Generating Custom Pooled CRISPR Libraries for Genetic Dissection of Biological Pathways.
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生成用于生物途径遗传解析的定制 CRISPR 文库。
DOI：
10.1007/978-1-0716-2209-4_21
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Gulbranson,DanielR
中科院分区：
文献类型：
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作者：
Gulbranson,DanielR
遗传筛选是解剖包括膜运输在内的生物途径的经典方法。聚集规律间隔短回文重复序列(CRISPR)/Cas9使这种方法在二倍体模型（包括培养的哺乳动物细胞）中得以应用。在这里，我们提出了生成自定义CRISPR库的详细协议。这些方法可用于为缺乏现有全基因组文库的新模式生物生成全基因组文库，也可用于生成较小的集中文库。
Genetic screens are a classic approach to dissecting biological pathways including membrane trafficking. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 have enabled the utility of this approach in diploid models, including cultured mammalian cells. Here, we present detailed protocols for generating custom CRISPR libraries. These methods are useful for generating genome-wide libraries for new model organisms that lack an existing genome-wide library, and for generating smaller focused libraries.