使用慢病毒CRISPR/Cas9系统在髓系细胞系中产生基因敲除
Generation of Genetic Knockouts in Myeloid Cell Lines Using a Lentiviral CRISPR/Cas9 System.
作者信息
Baker Paul J, Masters Seth L
机构信息
Inflammation division, The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, VIC, 3052, Australia.
Department of Medical Biology, The University of Melbourne, Parkville, VIC, 3010, Australia.
出版信息
Methods Mol Biol. 2018;1714:41-55. doi: 10.1007/978-1-4939-7519-8_3.
CRISPR/Cas9-based gene targeting allows deletion of a gene of interest from cultured cell lines. Due to difficulty in transiently transfecting hematopoetic cells with components required for this process, we have adopted a lentiviral system for delivery of the CRISPR/Cas9 components into myeloid cell lines. Here, we detail the process of knocking out genes from pools of cultured myeloid cells using this CRISPR/Cas9 system and describe methods of validating these knockout pools.
基于CRISPR/Cas9的基因靶向技术可从培养的细胞系中删除感兴趣的基因。由于难以用该过程所需的成分瞬时转染造血细胞，我们采用了慢病毒系统将CRISPR/Cas9成分导入髓系细胞系。在此，我们详细介绍了使用该CRISPR/Cas9系统从培养的髓系细胞库中敲除基因的过程，并描述了验证这些基因敲除库的方法。
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