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Strategies for the Enrichment and Selection of Genetically Modified Cells

Author:
Ren, Chonghua, Xu, Kun, Segal, David Jay, Zhang, Zhiying
Source:
Trends in biotechnology 2019 v.37 no.1 pp. 56-71
ISSN:
0167-7799
Subject:
gene editing, gene therapy, genome, genomics, loci, nucleases, screening
Abstract:
Programmable artificial nucleases have transitioned over the past decade from ZFNs and TALENs to CRISPR/Cas systems, which have been ubiquitously used with great success to modify genomes. The efficiencies of knockout and knockin vary widely among distinct cell types and genomic loci and depend on the nuclease delivery and cleavage efficiencies. Moreover, genetically modified cells are almost phenotypically indistinguishable from normal counterparts, making screening and isolating positive cells rather challenging and time-consuming. To address this issue, we review several strategies for the enrichment and selection of genetically modified cells, including transfection-positive selection, nuclease-positive selection, genome-targeted positive selection, and knockin-positive selection, to provide a reference for future genome research and gene therapy studies.