Double strand interaction is the predominant pathway for intermolecular recombination of adeno-associated viral genomes.
Intermolecular recombination is the foundation for dual vector mediated larger gene transfer by recombinant adeno-associated virus (rAAV). To identify precursors for intermolecular recombination, we sequentially infected skeletal muscle with AAV LacZ trans-splicing viruses. At 1 month postinfection, nearly all inputting single-strand (ss) AAV genomes were cleared out in muscle. If ss-ss interaction is absolutely required for intermolecular recombination, LacZ expression from sequential infection will be negligible to that from coinfection. Interestingly, expression from sequential infection reached approximately 50% of that from coinfection at the 1-month time-point in BL6 mice. In immune deficient SCID mice, expression from sequential infection was comparable to that from coinfection at the 4- and 13-month time points. Our results suggest that ds interaction represents the predominant pathway for AAV intermolecular recombination.
Yue
Yongping
Y
Department of Molecular Microbiology and Immunology, University of Missouri School of Medicine, Columbia, MO 65212, USA.
Duan
Dongsheng
D
D000818:Animals; D015139:Blotting, Southern; D004270:DNA, Circular / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D016679:Genome, Viral; D007763:Lac Operon / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D018482:Muscle, Skeletal / Q000378:metabolism / Q000821:virology; D011995:Recombination, Genetic; D013997:Time Factors; D020040:Trans-Splicing
D004270:DNA, Circular; D004279:DNA, Viral