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用于“鼠源化”人类细胞的嗜亲性假型慢病毒转导的方案,同时降低实验室人员的感染风险。

Protocol for ecotropic pseudotyped lentiviral transduction of "murinized" human cells while decreasing risk of infection for laboratory personnel.

作者信息

Weber Sandra L K, Heinzel Thorsten, Iyer-Bierhoff Aishwarya

机构信息

Institute of Biochemistry and Biophysics, Centre for Molecular Biomedicine (CMB), Friedrich Schiller University, Hans-Knoell-Strasse 2, 07745 Jena, Germany.

Institute of Biochemistry and Biophysics, Centre for Molecular Biomedicine (CMB), Friedrich Schiller University, Hans-Knoell-Strasse 2, 07745 Jena, Germany.

出版信息

STAR Protoc. 2025 Jun 20;6(2):103708. doi: 10.1016/j.xpro.2025.103708. Epub 2025 Mar 21.

Abstract

Lentiviral systems of transduction are valuable tools to stably express transgenes of interest in human cell lines. However, their use risks infection of the lab workers. Here, we present a protocol for lentiviral vector system pseudotyping with the murine ecotropic envelope. We detail steps for producing ecotropic pseudotyped lentiviral particles and transiently expressing the Slc7a1 receptor for murine leukemia virus in target human cells. We then describe procedures for puromycin selection and the characterization of stable cell lines for transgene expression. For complete details on the use and execution of this protocol, please refer to Iyer-Bierhoff et al..

摘要

慢病毒转导系统是在人类细胞系中稳定表达感兴趣的转基因的宝贵工具。然而,使用它们存在感染实验室工作人员的风险。在此,我们展示了一种用鼠嗜亲性包膜对慢病毒载体系统进行假型化的方案。我们详细介绍了产生嗜亲性假型化慢病毒颗粒以及在目标人类细胞中瞬时表达鼠白血病病毒的Slc7a1受体的步骤。然后我们描述了嘌呤霉素筛选程序以及转基因表达稳定细胞系的表征。有关此方案的使用和执行的完整详细信息,请参考Iyer - Bierhoff等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/00e6/11981732/514861aa7faa/fx1.jpg

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