Strategies for the Enrichment and Selection of Genetically Modified Cells

Chonghua Ren; Kun Xu; David Jay Segal; Zhiying Zhang

doi:10.1016/j.tibtech.2018.07.017

Strategies for the Enrichment and Selection of Genetically Modified Cells

Trends Biotechnol. 2019 Jan;37(1):56-71. doi: 10.1016/j.tibtech.2018.07.017. Epub 2018 Aug 19.

Authors

Chonghua Ren¹, Kun Xu², David Jay Segal³, Zhiying Zhang⁴

Affiliations

¹ School of Life Sciences, South China Normal University, Guangzhou, 510631, China; College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China; Genome Center and Department of Biochemistry and Molecular Medicine, University of California, Davis, CA 95616, USA; These authors contributed equally to this article.
² College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China; These authors contributed equally to this article.
³ Genome Center and Department of Biochemistry and Molecular Medicine, University of California, Davis, CA 95616, USA.
⁴ College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China. Electronic address: zhangzhy@nwsuaf.edu.cn.

PMID: 30135027
DOI: 10.1016/j.tibtech.2018.07.017

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.

Keywords: Artificial nucleases; enrichment; genetically modified cells; genome editing; selection.

Publication types

Research Support, Non-U.S. Gov't
Review

MeSH terms

Cell Separation / methods*
Gene Editing / methods*
Selection, Genetic*
Staining and Labeling / methods*