Viral Packaging and Transduction of Adult Hippocampal Neural Progenitors

Department of Chemical Engineering, University of California, Berkeley, CA, USA.
Methods in molecular biology (Clifton, N.J.) (Impact Factor: 1.29). 01/2010; 621:103-16. DOI: 10.1007/978-1-60761-063-2_7
Source: PubMed


Genetic manipulation of adult hippocampal neural progenitor cells is a useful technique for exploring gene function through gain of function and loss of function mutations or RNAi. Furthermore, the introduction of new genes can "re-program" progenitor cell behavior to force a desired lineage in signaling environments that are not normally permissive for that cell fate. Additionally, by using a systems biology approach, neural progenitors can even be taught new behaviors and responses to signaling. In this chapter, we describe protocols for retroviral and lentiviral packaging and transduction of progenitors. These techniques are important for studying the role of various genes in progenitor fate choice.

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Available from: David Vernon Schaffer
    • "The RAW/LR5 Lifeact-mCherry cell line was created by retroviral transduction using the vector pTK93_Lifeact- mCherry (Addgene plasmid 46357), which was generously deposited by Dr. Iain Cheeseman. Retrovirus was packaged using 293T cells, purified by ultracentrifugation , and titered by flow cytometry as previously described (Peltier and Schaffer, 2010). RAW/LR5 cells were infected at a multiplicity of infection of 0.25 and treated with 3 lg/mL puromycin for 6 days to select transduced cells. "
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