Optimization of vesicular stomatitis virus-G pseudotyped feline immunodeficiency virus vector for minimized cytotoxicity with efficient gene transfer.
FIV-based lentiviral vector has shown a unique opportunity as an efficient gene delivery vehicle, especially to nondividing human cells. Here, we genetically reconstructed the FIV-based vector by serially deleting residual virus genes of gag and vif, leading to minimized cytotoxicity together with efficient virus production and gene transfer. The modified FIV- based vector was generated by transiently transfecting 293T cells with three plasmids of the gene transfer vector with minimal gag region, the packaging plasmid without vif and the VSV-G-expressing plasmid. The vector was routinely generated as many as 1 x 10(7) transducing particles per ml and easily concentrated by simple centrifugation. The cytotoxic effect significantly decreased in sensitive cells to FIV infection even at high multiplicity of infection (MOI), such as 500. Moreover, the transduction efficiency was consistently retained after cell cycle was arrested in a variety of human cells. Taken together, our results suggest that the modified VSV-G pseudotyped FIV-based vector efficiently transduce dividing and nondividing human cells with minimal cytotoxicity.
Institute for Cancer Research, Yonsei University College of Medicine, Seoul, South Korea.
Jae Jin
JJ
Song
Boyoung
B
Lee
Jin Woo
JW
Chang
Joo-Hang
JH
Kim
Yunhee Kim
YK
Kwon
Heuiran
H
Lee
D000818:Animals; D002415:Cats; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics / Q000502:physiology*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D016089:Immunodeficiency Virus, Feline / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000502:physiology*; D014721:Vesicular stomatitis Indiana virus / Q000145:classification / Q000235:genetics / Q000502:physiology*; D014759:Viral Envelope Proteins / Q000502:physiology*
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins