一种新颖的“底盘”PSC线，通过选择辅助RMCE进行精确的安全港站点集成

IF 0.8 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Stem cell research Pub Date : 2025-06-24 DOI:10.1016/j.scr.2025.103758

Merve Ay , Desi Veleva , Mohammad M. Chowdhury , Andrew B.J. Prowse , Dmitry A. Ovchinnikov

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引用次数: 0

摘要

我们在此描述了一种人类多能干细胞系（hPSC），设计用于作为受体（“底盘”细胞系），用于重组介导的盒式交换（RMCE）介导的DNA片段精确插入到安全港位点。为了使用Flp重组酶进行定向插入，我们使用可切除的阴性选择盒，以FRT和FRT3位点为侧翼，靶向AAVS1安全港等位基因之一。所获得的hPSC细胞系表现出典型的群体形态、多能性特征和正常的核型。我们期望它能为FRT/ frt3侧翼转基因提供一个有用的AAVS1安全港“着陆点”。

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A novel “chassis” PSC line for precise safe harbor site integration via selection-aided RMCE

We describe here a human pluripotent stem cell line (hPSC) designed to serve as an acceptor (a “chassis” cell line) for the recombination-mediated cassette exchange (RMCE)-mediated precise insertion of DNA fragments into a safe harbor locus. To enable the use of Flp recombinase for directional insertion, we targeted one of the AAVS1 safe harbor alleles with an excisable negative selection cassette flanked with FRT and FRT3 sites. The generated hPSC line displayed typical colony morphology, pluripotency signatures and normal karyotype. We expect it to provide a useful AAVS1 safe harbor “landing site” for FRT/FRT3-flanked transgenes.

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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.