Abstract
The ability to introduce DNA elements into host cells and analyze the effects has revolutionized modern biology. Here we describe a protocol to generate Moloney murine leukemia virus (MMLV)-based, replication-incompetent pseudotyped retrovirus capable of infecting axolotls and incorporating genetic information into their genome. When pseudotyped with vesicular stomatitis virus (VSV)-G glycoprotein, the retroviruses can infect a broad range of proliferative axolotl cell types. However, if the retrovirus is pseudotyped with an avian sarcoma leukosis virus (ASLV)-A envelope protein, only axolotl cells experimentally manipulated to express the cognate tumor virus A (TVA) receptor can be targeted by infections. These strategies enable robust transgene expression over many cell divisions, cell lineage tracing, and cell subtype targeting for gene expression.
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Acknowledgements
This work was supported by start-up funds from Brigham and Women’s Hospital (J.L.W.). The authors would like to thank Christina DeMaso in Dr. Connie Cepko’s lab (Harvard Medical School) for sharing similar protocols used in infecting mammalian cells.
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Kuo, TH., Whited, J.L. (2015). Pseudotyped Retroviruses for Infecting Axolotl. In: Kumar, A., Simon, A. (eds) Salamanders in Regeneration Research. Methods in Molecular Biology, vol 1290. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2495-0_10
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DOI: https://doi.org/10.1007/978-1-4939-2495-0_10
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