Chen Yangcan, Luo Shengqiu, Hu Yanping, Zhou Qi, Li Wei
Key Laboratory of Organ Regeneration and Reconstruction, State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China; Bejing Institute for Stem Cell and Regenerative Medicine, Beijing 100101, China.
Key Laboratory of Organ Regeneration and Reconstruction, State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
STAR Protoc. 2025 Mar 21;6(1):103538. doi: 10.1016/j.xpro.2024.103538. Epub 2025 Jan 6.
R2 retrotransposons can be harnessed to insert genes at targeted sites by all-RNA delivery, presenting a new technology for next-generation biotherapeutics. Here, we report a protocol for evaluating the gene integration activity of R2 retrotransposons in mammalian cells. We describe the construction of vectors separately expressing R2 protein and donor, the process of liposome transfection, and flow cytometry. This protocol provides a useful reporter system, which can be applied to evaluate the activity of other new retrotransposon systems. For complete details on the use and execution of this protocol, please refer to Chen et al..
R2逆转座子可通过全RNA递送用于在靶向位点插入基因，为下一代生物疗法提供了一项新技术。在此，我们报告了一种评估R2逆转座子在哺乳动物细胞中基因整合活性的方案。我们描述了分别表达R2蛋白和供体的载体构建、脂质体转染过程以及流式细胞术。该方案提供了一个有用的报告系统，可用于评估其他新型逆转座子系统的活性。有关本方案使用和实施的完整详细信息，请参阅Chen等人的研究。
