PMID: 15335219Sep 1, 2004Paper

Transduction of multiple cell types using improved conditions for gene delivery and expression of SV40 pseudovirions packaged in vitro

BioTechniques
Chava Kimchi-SarfatyMichael M Gottesman

Abstract

This comprehensive study demonstrates highly efficient transduction of a wide variety of human, murine, and monkey cell lines, using a procedure for in vitro packaging of plasmid DNA in recombinant simian virus 40 (SV40) capsid proteins to form pseudovirions. The pseudovirions are encapsidated by the VP1 major capsid protein, with no SV40 sequence requirement, and are able to carry up to 17.7 kb of supercoiled plasmid DNA. We developed a procedure to scale-up production of SV40 pseudovirions, as well as an efficient protocol to concentrate the virions with no loss of activity. We also developed a method that allows transduction of 10 times more cells than the original protocol. This protocol was tested using supercoiled in vitro-packaged plasmid carrying the human multidrug-resistance gene (MDR1 encoding P-glycoprotein; P-gp), or the enhanced green fluorescent protein reporter gene (EGFP) in .45 human lymphoblastoid cells and in K562 human erythroleukemia cells. Multiple transductions at 24-h intervals were shown to increase expression using the EGFP reporter gene. The protocols developed in this study establish in vitro-packaged SV40 pseudovirions as one of the most efficient gene delivery systems.

References

Oct 21, 1999·Journal of Cellular Physiology·D S Strayer
Jan 29, 2002·Human Gene Therapy·Chava Kimchi-SarfatyMichael M Gottesman
Mar 5, 2003·Human Gene Therapy·Chava Kimchi-SarfatyMichael M Gottesman
Mar 7, 2003·Gene Therapy·P CordelierD S Strayer

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Citations

Sep 16, 2005·Pigment Cell Research·Werner BerensJo Lambert
Sep 10, 2005·Human Gene Therapy·Chava Kimchi-SarfatyMichael M Gottesman
Feb 25, 2006·Cancer Gene Therapy·C Kimchi-SarfatyM M Gottesman
Apr 26, 2008·Pancreas·Nilly ShimonyYosef S Haviv
Jan 21, 2006·Cancer Gene Therapy·G R Devi
Jul 23, 2005·Cancer Gene Therapy·Tulin Budak-AlpdoganJoseph R Bertino
Jun 5, 2020·Science Advances·Mariska G M van RosmalenGijs J L Wuite

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