Polyethylenimine-based transfection method as a simple and effective way to produce recombinant lentiviral vectors.
HIV-1-derived lentiviral vectors (LvV) are within the most attractive gene delivery vehicles in the context of both dividing and quiescent cells. LvV is currently produced by the conventional calcium phosphate precipitation method. Nevertheless, this procedure is highly susceptible to variations in pH and impurities, which lead to inconsistencies in LvV production. Here, we present a simple and robust procedure for LvV production using branched 25 kDa polyethylenimine, with a transfection efficiency of over 90% and viral titer yields of about 1 x 10(7) infective lentiviral particles per milliliter. The procedure outlined is simple, consistent, and as inexpensive as the CaPO(4)-based method.
Toledo
Jorge R
JR
Animal Biotechnology Department, Centre for Genetic Engineering and Biotechnology, Havana, Cuba.
Prieto
Yanet
Y
Oramas
Nayrobis
N
Sánchez
Oliberto
O
D002460:Cell Line; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D011094:Polyethyleneimine; D014162:Transfection / Q000379:methods*
D011094:Polyethyleneimine