Fusion-mediated injection of SV40-DNA. Introduction of SV40-DNA into tissue culture cells by the use of DNA-loaded reconstituted Sendai virus envelopes

A. Loyter*, A. Vainstein, M. Graessmann, A. Graessmann

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

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 giycoproteins 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.

Original languageAmerican English
Pages (from-to)415-425
Number of pages11
JournalExperimental Cell Research
Volume143
Issue number2
DOIs
StatePublished - Feb 1983

Bibliographical note

Funding Information:
This work was supported by a grant from the Deutsche Forschungsgemeinschaft no. GR 384-S (to A. G.) and from the March of Dimes Birth Defects Foundation;

Funding Information:
Part of this work was performed during a visit of A. V. to the Free University of Berlin, which was supported by a short-term fellowship from EMBO, European Molecular Biology Organization.

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