... Protocols Lentivirus Production Lentivirus Production You may also ...for alternative packaging cell lines or transfection reagents. Once produced, lentivirus can be used for... Timeline Day 0: Seed 293T packaging cells Day 1 (pm): Transfect packaging cells Day 2 (am): 18 h post-transfection...this protocol to generate lentivirus Protocols...Introduction This protocol can be used to produce lentivirus from a lentiviral vector transfected into 293T...Considerations Before You Start The health of the packaging cell line is critical for obtaining high viral... 15 for viral production. Procedure Seed 293T packaging cells at 3.8×10 6 cells per plate in DMEM Complete...

...control vector containing scrambled shRNA. psPAX2 Packaging plasmid for producing viral particles. pMD2.G ...particles. Note: pLKO.1 can also be used with packaging plasmid pCMV-dR8.2 dvpr and envelope plasmid pCMV-VSVG...tubes VWR: #87003-290 Note: pLKO.1 could also be packaged using pCMV-dR8.2 dvpr and pCMV-VSVG from the Robert...transfection: 1 μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging plasmid 250 ng pMD2.G envelope plasmid to 20 μl... may want to vary the ratio of shRNA plasmid, packaging plasmid, and envelope plasmid to obtain the ratio...DS et. al. Cell . 2003. 115:199-208. PubMed . Lentivirus-delivered stable gene silencing by RNAi in primary...1997. 15(9):871-5. PubMed . Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery...