Forced recombination of psi-modified murine leukaemia virus-based vectors with murine leukaemia-like and VL30 murine endogenous retroviruses.
Co-encapsidation of retroviral RNAs into virus particles allows for the generation of recombinant proviruses through events of template switching during reverse transcription. By use of a forced recombination system based on recombinational rescue of replication- defective primer binding site-impaired Akv-MLV-derived vectors, we here examine putative genetic interactions between vector RNAs and copackaged endogenous retroviral RNAs of the murine leukaemia virus (MLV) and VL30 retroelement families. We show (i) that MLV recombination is not blocked by nonhomology within the 5' untranslated region harbouring the supposed RNA dimer-forming cis -elements and (ii) that copackaged retroviral RNAs can recombine despite pronounced sequence dissimilarity at the cross-over site(s) and within parts of the genome involved in RNA dimerization, encapsidation and strand transferring during reverse transcription. We note that recombination-based rescue of primer binding site knock-out retroviral vectors may constitute a sensitive assay to register putative genetic interactions involving endogenous retroviral RNAs present in cells of various species.
Mikkelsen
Jacob Giehm
JG
Department of Molecular and Structural Biology1 and Department of Medical Microbiology and Immunology,2 University of Aarhus, C. F. Moellers Allé, Bldg 130, DK-8000 Aarhus, Denmark.
Lund
Anders H
AH
Department of Molecular and Structural Biology1 and Department of Medical Microbiology and Immunology,2 University of Aarhus, C. F. Moellers Allé, Bldg 130, DK-8000 Aarhus, Denmark.
Duch
Mogens
M
Department of Molecular and Structural Biology1 and Department of Medical Microbiology and Immunology,2 University of Aarhus, C. F. Moellers Allé, Bldg 130, DK-8000 Aarhus, Denmark.
Pedersen
Finn Skou
FS
Department of Molecular and Structural Biology1 and Department of Medical Microbiology and Immunology,2 University of Aarhus, C. F. Moellers Allé, Bldg 130, DK-8000 Aarhus, Denmark.
D020121:5' Untranslated Regions; D001483:Base Sequence; D020077:Endogenous Retroviruses / Q000235:genetics*; D005822:Genetic Vectors; D009052:Leukemia Virus, Murine / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D020079:Terminal Repeat Sequences; D019065:Virus Assembly
D020121:5' Untranslated Regions