Generation of stable Cas9-EGFP expressing human induced pluripotent stem cell lines based on SeLection by Essential-gene Exon Knock-in technology

Here, we used SeLection by Essential-gene Exon Knock-in technology to generate the iPSC line with constitutive expression of Cas9-EGFP, while retaining all functions of the essential gene. Cas9-EGFP was inserted into the GAPDH exon9 via the homologous recombination, avoiding Cas9 silencing that often occurs during iPSC differentiation. The edited cell line shows precise knock-in locus with the typical characteristics and pluripotency of iPSCs. Therefore, this iPSC line is valuable for CRISPR screening or related experiments and could be widely used in the CRISPR/Cas9-based gene editing.

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Publication Details

Journal
Stem Cell Research
Published
2025-04-08
DOI
https://doi.org/10.1016/j.scr.2025.103710
Citations
1
Primary Topic
CRISPR and Genetic Engineering
Type
article
Field-Weighted Citation Impact
0.81
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article

Generation of stable Cas9-EGFP expressing human induced pluripotent stem cell lines based on SeLection by Essential-gene Exon Knock-in technology

Joe Z. Zhang, E. V. Zaklyazminskaya, Hao Yang, Lokyu Cheng et al.
1 citations
Stem Cell Research
CRISPR and Genetic Engineering
0.81
article

Generation of stable Cas9-EGFP expressing human induced pluripotent stem cell lines based on SeLection by Essential-gene Exon Knock-in technology

Joe Z. Zhang, E. V. Zaklyazminskaya, Hao Yang, Lokyu Cheng, Olga S. Sokolova, Yao Zhang, Zijun Lu, Hongmei Tan
article en
1 citations

Abstract

Here, we used SeLection by Essential-gene Exon Knock-in technology to generate the iPSC line with constitutive expression of Cas9-EGFP, while retaining all functions of the essential gene. Cas9-EGFP was inserted into the GAPDH exon9 via the homologous recombination, avoiding Cas9 silencing that often occurs during iPSC differentiation. The edited cell line shows precise knock-in locus with the typical characteristics and pluripotency of iPSCs. Therefore, this iPSC line is valuable for CRISPR screening or related experiments and could be widely used in the CRISPR/Cas9-based gene editing.

Stem Cell ResearchVol. 85
Sun Yat-sen University (CN), Harbin Institute of Technology (CN), The Seventh Affiliated Hospital of Sun Yat-sen University (CN), Shenzhen Bay Laboratory (CN), Shenzhen Technology University (CN), Russian Scientific Center of Surgery (RU), Shenzhen MSU-BIT University, University of Hong Kong (HK)
Zero hunger
Openalex Percentile: Top 29%
CRISPR and Genetic Engineering
0.81
DOIOpen Access PDFOpenAlex
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Generation of stable Cas9-EGFP expressing human induced pluripotent stem cell lines based on SeLection by Essential-gene Exon Knock-in technology — Joe Z. Zhang, E. V. Zaklyazminskaya, et al. · Stem Cell Research (2025) | TGRS Research Map | TGRS