TRIM5α/Cyclophilin A-Modified MDBK Cells for Lentiviral-Based Gene Editing

文献类型： 外文期刊

第一作者： Wo, Lijing

作者： Wo, Lijing;Qi, Shuhui;Guo, Yongqi;Sun, Chao;Yin, Xin

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关键词： MDBK; TRIM5 alpha; CypA

期刊名称：VIRUSES-BASEL （ 影响因子：3.5； 五年影响因子：3.7 ）

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年卷期： 2025 年 17 卷 7 期

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收录情况： SCI

摘要： The human immunodeficiency virus 1 (HIV-1)-based lentivirus has been widely used for genetic modification. However, the efficiency of lentiviral-based gene modification in Madin-Darby bovine kidney (MDBK) cells is considerably limited. In this study, we have shown that siRNA-mediated depletion of TRIM5 alpha, a restriction factor in HIV-1 infection, can dramatically enhance HIV-1 infection in MDBK cells. Furthermore, we generated a doxycycline-inducible Cas9-overexpressing MDBK cell line (MDBK-iCas9) suitable for CRISPR/Cas9-mediated editing. On this basis, we created a TRIM5 alpha knock-out MDBK-iCas9 cell line MDBK-iCas9TRIM5 alpha-/- without additional genome insertions by combining sgRNA transfection and single-cell cloning. We found that MDBK-iCas9TRIM5 alpha-/- displayed greater permissiveness to lentivirus infection compared with MDBK-WT cells. Notably, we found that treatment with the chemical compound cyclosporine A, which directly interacts with cell factor cyclophilin A (CypA), could markedly increase the infectivity of lentivirus in both MDBK-iCas9TRIM5 alpha-/- and MDBK-WT cell lines, suggesting that CypA functions independently with TRIM5 alpha as an inhibitor of the lentivirus in bovine cells. Therefore, combining bovine TRIM5 alpha and CypA targeting could remarkably enhance lentivirus infection. In conclusion, our findings highlight a promising gene engineering strategy for bovine cells that can surmount the significant barriers to investigating the interplay between bovine viruses and their host cells.

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