Gene delivery via polyomavirus major capsid protein VP(1), isolated from recombinant Escherichia coli.
In this study we attempted to investigate the feasibility of transferring exogenous DNA into the recipient host via polyomavirus major capsid protein VP(1) pseudocapsids generated from recombinant Escherichia coli, designated PyVP(1,E), both in vitro and in vivo. NIH-3T3 and FR3T3 cells were transfected with pCMV beta and pPyMT-1 plasmid DNA, respectively. In vitro DNA transfection was carried out via Pseudofect- and PyVP(1,E)-mediated methods, or by co-precipitation with calcium phosphate. Expression of beta-galactosidase and PyMT reporter genes was examined by Western-blot analysis. Parallel experiments were performed in vivo by direct injection of pCMV beta and pPyMT-1 plasmid DNA, complexed with PyVP(1,E), into the livers of Wistar rats, followed by Western-blot analysis and histochemical staining. The results obtained from in vitro transfection experiments showed that expression of the reporter genes can be detected in the recipient cells at 48 h post-transfection. PyVP(1,E) was shown to exhibit similarly efficient in vitro DNA transfection properties to Pseudofect, which was obtained from recombinant baculovirus.
School of Pharmacy, College of Medicine, National Taiwan University, 1, Jen-Ai Road, Section 1, Taipei 100, Taiwan.ywyang@ha.mc.ntu.edu.tw
Y W
YW
Yang
L H
LH
Chen
D016475:3T3 Cells; D000818:Animals; D019556:COS Cells; D002213:Capsid / Q000008:administration & dosage / Q000235:genetics* / Q000302:isolation & purification; D036022:Capsid Proteins; D004926:Escherichia coli / Q000235:genetics*; D018014:Gene Transfer Techniques; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D011107:Polylysine / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics / Q000302:isolation & purification; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D036022:Capsid Proteins; D011994:Recombinant Proteins; C062481:VP1 protein, polyomavirus; D011107:Polylysine; D001616:beta-Galactosidase