TY - JOUR
T1 - Strategies for the Enrichment and Selection of Genetically Modified Cells
AU - Ren, Chonghua
AU - Xu, Kun
AU - Segal, David
AU - Zhang, Zhiying
PY - 2018/1/1
Y1 - 2018/1/1
N2 - 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.
AB - 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.
KW - Artificial nucleases
KW - enrichment
KW - genetically modified cells
KW - genome editing
KW - selection
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U2 - 10.1016/j.tibtech.2018.07.017
DO - 10.1016/j.tibtech.2018.07.017
M3 - Article
C2 - 30135027
AN - SCOPUS:85051650599
JO - Trends in Biotechnology
JF - Trends in Biotechnology
SN - 0167-7799
ER -