Protocol for evaluating the activity of R2 retrotransposons in mammalian cells
Summary: 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 con...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724007032 |
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| author | Yangcan Chen Shengqiu Luo Yanping Hu Qi Zhou Wei Li |
| author_facet | Yangcan Chen Shengqiu Luo Yanping Hu Qi Zhou Wei Li |
| author_sort | Yangcan Chen |
| collection | DOAJ |
| description | Summary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-6bd81c9db37747819f7c79313b71eb0d |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-6bd81c9db37747819f7c79313b71eb0d2025-01-07T04:17:30ZengElsevierSTAR Protocols2666-16672025-03-0161103538Protocol for evaluating the activity of R2 retrotransposons in mammalian cellsYangcan Chen0Shengqiu Luo1Yanping Hu2Qi Zhou3Wei Li4Key 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; Corresponding authorKey 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, ChinaKey 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, ChinaKey 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; Bejing Institute for Stem Cell and Regenerative Medicine, Beijing 100101, China; Corresponding authorKey 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; Bejing Institute for Stem Cell and Regenerative Medicine, Beijing 100101, China; Corresponding authorSummary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724007032molecular biologyCRISPRbiotechnology and bioengineering |
| spellingShingle | Yangcan Chen Shengqiu Luo Yanping Hu Qi Zhou Wei Li Protocol for evaluating the activity of R2 retrotransposons in mammalian cells STAR Protocols molecular biology CRISPR biotechnology and bioengineering |
| title | Protocol for evaluating the activity of R2 retrotransposons in mammalian cells |
| title_full | Protocol for evaluating the activity of R2 retrotransposons in mammalian cells |
| title_fullStr | Protocol for evaluating the activity of R2 retrotransposons in mammalian cells |
| title_full_unstemmed | Protocol for evaluating the activity of R2 retrotransposons in mammalian cells |
| title_short | Protocol for evaluating the activity of R2 retrotransposons in mammalian cells |
| title_sort | protocol for evaluating the activity of r2 retrotransposons in mammalian cells |
| topic | molecular biology CRISPR biotechnology and bioengineering |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724007032 |
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