Development of packaging cell lines for generation of adeno-associated virus vectors by lentiviral gene transfer of trans-complementary components

European Journal of Haematology
Satoki NakamuraYuichi Iwaki

Abstract

Adeno-associated virus (AAV) vector system has several useful advantages with regard to in vitro and in vivo gene transfer. However, their usages have been limited by cumbersome and labor-intensive vector production in the traditional method. To overcome limitations in AAV production, in this report, we explored the possibility of generating AAV packaging cell line, 293T R/C.VA.E2A.E4. cells, by using lentivirus-mediated transduction of Rep/Cap gene of AAV-2, VA RNA, E2A, and E4 genes of Ad5 into 293T cells. In packaging cell lines, it is important that supply of the AAV vector can be stably performed for long time. We showed that the 293T R/C.VA.E2A.E4. cells have stably maintained the transduced components after more than 10 passages and yielded high-titer AAV vectors, and the titer of AAV vectors did not decline even if culture of the packaging cells was continued for long time. The Rep/Cap and E4 gene products caused no remarkable cytotoxicity. The 293T R/C.VA.E2A.E4. cells might be able to tolerate the Rep/Cap and E4 gene products, or have less copy numbers of the Rep/Cap and E4 genes than the traditional method. Moreover, we showed that the AAV vectors derived from 293T R/C.VA.E2A.E4. cells infected the primary human CD34...Continue Reading

References

Oct 1, 1992·Journal of Virology·Q YangJ P Trempe
Oct 1, 1994·Molecular Biotechnology·D S Latchman
Nov 27, 1995·Annals of the New York Academy of Sciences·K I Berns, C Giraud
Oct 1, 1995·Human Gene Therapy·K R ClarkP R Johnson
Feb 18, 1997·Proceedings of the National Academy of Sciences of the United States of America·M NevelsT Dobner
Apr 29, 1998·Human Gene Therapy·A SalvettiP Moullier
Oct 10, 1998·Journal of Virology·T DullL Naldini
Nov 30, 1999·Molecular Cell Biology Research Communications : MCBRC·K W Adolph, P Bornstein

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Citations

May 27, 2005·Human Gene Therapy·S Zolotukhin
Nov 22, 2007·Experimental Neurology·Ayse UlusoyDeniz Kirik

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