基于关键基因外显子敲入技术的选择生成稳定表达Cas9-EGFP的人诱导多能干细胞系

IF 0.8 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Stem cell research Pub Date : 2025-04-07 DOI:10.1016/j.scr.2025.103710

Yao Zhang , Zijun Lu , Hao Yang , Lokyu Cheng , Elena Zaklyazminskaya , Olga Sokolova , Hongmei Tan , Joe Z Zhang

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引用次数: 0

摘要

在这里，我们使用SeLection by essential -gene Exon Knock-in技术生成了Cas9-EGFP组成表达的iPSC系，同时保留了必需基因的所有功能。Cas9- egfp通过同源重组插入GAPDH外显子9，避免了iPSC分化过程中经常发生的Cas9沉默。编辑后的细胞系显示出精确的敲入位点，具有iPSCs的典型特征和多能性。因此，该iPSC系具有CRISPR筛选或相关实验的价值，可广泛应用于基于CRISPR/ cas9的基因编辑。

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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

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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来源期刊

Stem cell research 生物-生物工程与应用微生物

CiteScore

2.20

自引率

8.30%

发文量

338

审稿时长

55 days

期刊介绍： Stem Cell Research is dedicated to publishing high-quality manuscripts focusing on the biology and applications of stem cell research. Submissions to Stem Cell Research, may cover all aspects of stem cells, including embryonic stem cells, tissue-specific stem cells, cancer stem cells, developmental studies, stem cell genomes, and translational research. Stem Cell Research publishes 6 issues a year.