Generation of Genetic Knockouts in Myeloid Cell Lines Using a Lentiviral CRISPR/Cas9 System
Generation of Genetic Knockouts in Myeloid Cell Lines Using a Lentiviral CRISPR/Cas9 System
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DOI：
10.1007/978-1-4939-7519-8_3
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发表时间：
2018-01-01
期刊：
影响因子：
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通讯作者：
Masters, Seth L.
中科院分区：
文献类型：
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作者：
Baker, Paul J.;Masters, Seth L.
基于CRISPR/Cas9的基因打靶允许从培养的细胞系中删除感兴趣的基因。由于难以瞬时地将CRISPR/Cas9组分转入造血细胞，我们采用了慢病毒系统将CRISPR/Cas9组分导入髓系细胞系。在这里，我们详细介绍了使用CRISPR/Cas9系统从培养的髓系细胞池中敲除基因的过程，并描述了验证这些敲除池的方法。
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.