2022•Unpublished venueOpen access

Generation of stable cell lines via retroviral or lentiviral transduction v1

Rotimi Yemi Fasimoye, Dario R. Alessi

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Abstract

The ability to stably express a protein of interest in cells is critical to study its function. Here, we describe a protocol to generate stable cell lines using a retrovirus system that can be used for a variety of mouse and human cell lines. Our protocol includes the production of retroviruses encoding the transgene of interest in HEK293FT and the subsequent transduction of the cell line that is intended to stably express the protein of interest. The same protocol can also be used to generate stable cell lines using a lentivirus system. It should be noted that when using this method, the transgene of interest will be randomly integrated into the cell genome.

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The ability to stably express a protein of interest in cells is critical to study its function. Here, we describe a protocol to generate stable cell lines using a retrovirus system that can be used for a variety of mouse and human cell lines. Our protocol includes the production of retroviruses encoding the transgene of interest in HEK293FT and the subsequent transduction of the cell line that is intended to stably express the protein of interest. The same protocol can also be used to generate stable cell lines using a lentivirus system. It should be noted that when using this method, the transgene of interest will be randomly integrated into the cell genome.

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Available abstract

The ability to stably express a protein of interest in cells is critical to study its function. Here, we describe a protocol to generate stable cell lines using a retrovirus system that can be used for a variety of mouse and human cell lines. Our protocol includes the production of retroviruses encoding the transgene of interest in HEK293FT and the subsequent transduction of the cell line that is intended to stably express the protein of interest. The same protocol can also be used to generate stable cell lines using a lentivirus system. It should be noted that when using this method, the transgene of interest will be randomly integrated into the cell genome.

Key concepts: Retrovirus, Transduction (biophysics), Transgene, Cell culture, Biology, Cell biology, Cell, Computational biology

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