How to produce or package recombinant lentiviruses?

The production of recombinant lentiviruses requires the packaging cell line (e.g., HEK293T), the transfer plasmid, and the packaging plasmids. The components necessary for the lentivirus production are split into the transfer and packaging plasmids to increase the safety. The transfer plasmid contains in-cis elements required for lentivirus production including two LTRs, the packaging signal , and the transgene expression cassette. The packaging plasmids contain in-trans elements required for lentivirus production including gal, pol, envelope (typically VSV-G). By co-transfection of the transfer and packaging plasmids into the packaging cell line, recombinant lentiviral particles are assembled within the packaging cells and are subsequently released into the supernatant of the culture. The supernatant containing the crude lentiviral particles can be harvested and further concentrated. Crude or concentrated virus can then be used to transduce the cells of interest, or stored at -80 ℃ as soon as possible to avoid loss of titer.

RGBiotech offers a full set of plasmids required for producing lentiviral particles. The standard target gene expression plasmid is pLVX-Puro. However, we can replace the pLVX-Puro plasmid with one fit-for-project plasmid according to different requirements.

For detailed lentivirus packaging protocol, please see our Lentivirus Production Guideline.

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