Homologous recombination of exogenous DNA fragments with genomic DNA in somatic cells of mice.
We compared liposomes and empty viral capsids for their use as vehicles for DNA transfer into cells and animals. DNA binding capacity was high for liposomes, but DNase I protection of DNA bound to liposomes was only moderate in comparison to DNA incorporated into viral capsids. Cellular uptake of radiolabeled and physiologically active DNA was also compared. For animal studies we chose an endogenous retroposon as target gene. To identify recombinational events we replaced a part of this gene with an artificial sequence not present in the mouse genome. The recombination rate for DNA fragments transfected in Polyoma capsids in live mice was higher than for liposome mediated transfection. Homologous recombination could be observed for both DNA transfer methods, mediated by positively charged liposomes (DOTMA) and by empty Polyoma viral capsids.
Gareis
M
M
Clin. Res. Units Rheumatol., Max-Planck Soc./University of Erlangen-Nürnberg, FRG.
Harrer
P
P
Bertling
W M
WM
D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000378:metabolism*; D002460:Cell Line; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors; D008081:Liposomes / Q000378:metabolism*; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D011120:Polyomavirus / Q000235:genetics; D011995:Recombination, Genetic; D014158:Transcription, Genetic; D014162:Transfection
D008081:Liposomes; D004247:DNA