Development of a universal antibiotic resistance screening system for efficient enrichment of C-to-G and A-to-G base editing

文献类型： 外文期刊

第一作者： Xin, Ying

作者： Xin, Ying;Lu, Hongjiang;Xin, Ying;Feng, Hu;He, Chenfei;Lu, Hongjiang;Zuo, Erwei;Yan, Nana

作者机构：

关键词： CGBE base editors; ABE base editors; Universal enrichment vector; Screening; Base-editing activity

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.2； 五年影响因子：7.8 ）

ISSN： 0141-8130

年卷期： 2024 年 268 卷

页码：

收录情况： SCI

摘要： To expand the scope of genomic editing, a C-to-G transversion-based editor called CGBE has been developed for precise single-nucleotide genomic editing. However, limited editing efficiency and product purity have hindered the development and application of CGBE. In this study, we introduced the Puromycin-Resistance Screening System, referred to as CGBE/ABE-PRSS, to select genetically modified cells via the CGBE or ABE editors. The CGBE/ABE-PRSS system significantly improves the enrichment efficiency of CGBE- or ABE-modified cells, showing enhancements of up to 59.6 % compared with the controls. Our findings indicate that the CGBE/ABEPRSS, when driven by the CMV promoter, results in a higher enrichment of edited cells compared to the CAG and EF1 alpha promoters. Furthermore, we demonstrate that this system is compatible with different versions of both CGBE and ABE, enabling various cell species and simultaneous multiplexed genome editing without any detectable random off-targets. In conclusion, our developed CGBE/ABE-PRSS system facilitates the selection of edited cells and holds promise in both basic engineering and gene therapy applications.

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