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. 2018:1714:41-55.
doi: 10.1007/978-1-4939-7519-8_3.

Generation of Genetic Knockouts in Myeloid Cell Lines Using a Lentiviral CRISPR/Cas9 System

Paul J Baker  1   2 , Seth L Masters  3   4
Affiliations

Generation of Genetic Knockouts in Myeloid Cell Lines Using a Lentiviral CRISPR/Cas9 System

Paul J Baker et al. Methods Mol Biol. 2018.

Abstract

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.

Keywords: CRISPR; CRISPR validation; Immortalized BMDM; Lentivirus; Myeloid; THP-1.

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