title: Gene targeting by adeno-associated virus vectors is cell-cycle dependent.
abstract: Adeno-associated virus (AAV) vectors can be used to introduce site-specific mutations into homologous chromosomal sequences. There are many potential applications of this technique, but the process of AAV-mediated gene targeting and factors that influence targeting efficiency are not completely understood. We investigated the dependence of AAV-mediated gene targeting on the host cell-cycle status. The frequency of gene targeting by AAV vectors was compared in dividing and serum-arrested normal human fibroblast cultures. Gene targeting occurred in arrested fibroblast cultures at 0.15 to 1.1% the frequency of dividing cultures, and only took place in cells that had undergone DNA synthesis. Gene targeting was also reduced when DNA synthesis was inhibited by hydroxyurea.
authors.affiliation: Division of Hematology, Department of Medicine, University of Washington, Seattle, WA 98195, USA.
authors.forename: Grant
authors.initials: G
authors.lastname: Trobridge
authors.forename: Roli K
authors.initials: RK
authors.lastname: Hirata
authors.forename: David W
authors.initials: DW
authors.lastname: Russell
mesh_terms: D001973:Bromodeoxyuridine / Q000378:metabolism; D002453:Cell Cycle / Q000235:genetics*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004247:DNA / Q000096:biosynthesis; D004261:DNA Replication; D000229:Dependovirus / Q000235:genetics*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D008297:Male
chemical_list: D016895:Culture Media, Serum-Free; D004247:DNA; D007041:Hypoxanthine Phosphoribosyltransferase; D001973:Bromodeoxyuridine; D006918:Hydroxyurea