Capsid-targeted viral inactivation can eliminate the production of infectious murine leukemia virus in vitro.
Capsid-targeted viral inactivation (CTVI), a promising gene-based antiviral strategy against retroviruses, was designed to disrupt the retroviral life cycle by incorporating a degradative enzyme (e.g., nuclease) into viral particles during assembly, thereby reducing or eliminating the production of infectious virus. The experimental system used to develop the CTVI strategy for retroviruses is designed to block the production of infectious Moloney murine leukemia virus (Mo-MLV). Two nucleases, Escherichia coli ribonulease HI and Staphylococcus nuclease, have been shown to be tolerated by the cell as Mo-MLV Gag-nuclease fusion polyproteins and still be active in the viral particles. The goal of this study was to determine what cellular and viral factors limit CTVI in cultured cells. The avian DF-1 cell line greatly expanded our ability to test the antiviral efficacy of CTVI in long-term assays and to determine the mechanism(s) of CTVI action. The CTVI antiviral effect is dependent on the level of Mo-MLV Gag-nuclease fusion polyprotein expressed. The Mo-MLV Gag-nuclease polyproteins produce a long-term prophylactic antiviral effect after a low- or high-dose Mo-MLV challenge. The Mo-MLV Gag-nuclease fusions have a significant therapeutic effect ( approximately 1000-fold) on the production of infectious Mo-MLV. The therapeutic CTVI effect can be improved by a second delivery of the CTVI fusion gene. Both the prophylactic and the therapeutic CTVI antiviral approaches can virtually eliminate the production of infectious Mo-MLV in vitro and are only limited by the number of cells in the population that do not express adequate levels of the CTVI fusion polyprotein.
VanBrocklin
M
M
Molecular Medicine Program, Mayo Clinic and Mayo Foundation, Rochester, Minnesota, 55905, USA.
Federspiel
M J
MJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000235:genetics*; D002460:Cell Line; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015750:Genes, gag; D005822:Genetic Vectors; D051379:Mice; D008836:Micrococcal Nuclease / Q000235:genetics; D008979:Moloney murine leukemia virus / Q000235:genetics* / Q000254:growth & development / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D016914:Ribonuclease H / Q000235:genetics; D014779:Virus Replication
D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins; D016914:Ribonuclease H; C078856:ribonuclease HI; D008836:Micrococcal Nuclease