Fusion-mediated injection of SV40-DNA. Introduction of SV40-DNA into tissue culture cells by the use of DNA-loaded reconstituted Sendai virus envelopes.
Sendai virus envelopes can be solubilized by non-ionic detergents such as Triton X-100. Removal of the detergent from a supernatant containing the solubilized viral envelope glycoproteins results in the formation of reconstituted fusogenic viral envelopes. When SV40-DNA is added to the reconstitution system, it is trapped within the viral envelope. Incubation of SV40-DNA-loaded Sendai virus envelopes with permissive cells (CV1 and TC7 cells) resulted in fusion-mediated injection of the trapped DNA, as was demonstrated by the ability of the injected cells to synthesize SV40-T-antigen. Quantitative estimation revealed that up to 20% of the injected cells were able to synthesize T-antigen. Loaded viral envelopes were able to inject SV40-DNA and to promote synthesis of T-antigen also in cells which are resistant to infection by intact SV40 viruses, such as F1' 1-4 cells. In addition, it is shown that reconstituted envelopes of Sendai virus are able to transfer membrane fragments from SV40 receptor-positive into SV40 receptor-negative cells, such as F1' 1-4 cells. After implantation of SV40 receptors, the F1' 1-4 cells became susceptible to infection by intact SV40 viruses.
D000818:Animals; D000956:Antigens, Viral; D000957:Antigens, Viral, Tumor; D002460:Cell Line; D002522:Chlorocebus aethiops; D004279:DNA, Viral; D008561:Membrane Fusion; D008845:Microinjections; D010222:Parainfluenza Virus 1, Human / Q000502:physiology*; D051381:Rats; D011991:Receptors, Virus / Q000378:metabolism; D013539:Simian virus 40 / Q000235:genetics* / Q000276:immunology; D014162:Transfection
D000956:Antigens, Viral; D000957:Antigens, Viral, Tumor; D004279:DNA, Viral; D011991:Receptors, Virus