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A Versatile Tool for the Quantification of CRISPR/Cas9-Induced Genome Editing Events in Human Hematopoietic Cell Lines and Hematopoietic Stem/Progenitor Cells

机译:用于量化人造血细胞系和造血干/祖细胞中CRISPR / CAS9诱导的基因组编辑事件的多功能工具

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摘要

The efficient site-specific DNA double-strand breaks (DSB) created by CRISPR/Cas9 has revolutionized genome engineering and has great potential for editing hematopoietic stem/progenitor cells (HSPCs). However, detailed understanding of the variables that influence choice of DNA-DSB repair (DDR) pathways by HSPC is required for therapeutic levels of editing in these clinically relevant cells. We developed a hematopoietic-reporter system that rapidly quantifies the three major DDR pathways utilized at the individual DSB created by CRISPR/Cas9-NHEJ, MMEJ, and HDR and show its applicability in evaluating the different DDR outcomes utilized by human hematopoietic cell lines and primary human HSPC. (C) 2018 Elsevier Ltd. All rights reserved.
机译:由CRISPR / CAS9产生的有效的特异性地点DNA双链断裂(DSB)具有革命性的基因组工程,并具有造成造血干/祖细胞(HSPC)的巨大潜力。 然而,对这些临床相关细胞中的编辑水平的治疗水平需要详细了解影响DNA-DSB修复(DDR)途径的选择的变量。 我们开发了一种造血记者系统,迅速量化了由CRISPR / CAS9-NHEJ,MMEJ和HDR创建的个体DSB中使用的三个主要DDR途径,并在评估人造血细胞系和初级利用的不同DDR结果方面表现出其适用性 人类HSPC。 (c)2018年elestvier有限公司保留所有权利。

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