title: Genome engineering using Adeno-Associated Virus (AAV).
abstract: The ability to edit the genome of cell lines has provided valuable insights into biological processes and the contribution of specific mutations to disease biology. These techniques fall into two categories based on the DNA repair mechanism that is used to incorporate the genetic change. Nuclease-based technologies, such as Zinc-Finger Nucleases, TALENS, and Crispr/Cas9, rely on non-homologous end-joining (NHEJ) and homology directed repair (HDR) to generate a range of genetic modifications. Adeno-Associated Virus (AAV) utilizes homologous recombination to generate precise and predictable genetic modifications directly at the target locus. AAV has been used to create over 500 human isogenic cell lines comprising a wide range of genetic alterations from gene knockouts, insertions of point mutations, indels, epitope tags, and reporter genes. Here we describe the generation and use of AAV gene targeting vectors and viruses to create targeted isogenic cell lines.
authors.affiliation: MedImmune, Milstein Building, Granta Park, Cambridge, CB21 6GH, UK, howesr@medimmune.com.
authors.forename: Rob
authors.initials: R
authors.lastname: Howes
authors.forename: Christine
authors.initials: C
authors.lastname: Schofield
mesh_terms: D018929:Cell Culture Techniques; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D018390:Gene Targeting; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics* / Q000302:isolation & purification; D005838:Genotype; D059765:Homologous Recombination; D006801:Humans; D014161:Transduction, Genetic