title: Combining CRISPR/Cas9 and rAAV Templates for Efficient Gene Editing.
abstract: Altering endogenous genes in cells is an integral tool of modern cell biology. The ease-of-use of the CRISPR/Cas9 system to introduce genomic DNA breaks at specific sites in vivo has led to its rapid and wide adoption. In the absence of a DNA template, the lesion is repaired by nonhomologous end joining resolving as internal deletions. However, in the presence of a homologous DNA template, homology-directed repair occurs with variable efficiencies. Recent work has demonstrated that highly efficient gene targeting can be induced by combining CRISPR/Cas9 targeting of genomic loci with recombinant adeno-associated virus (rAAV) to provide a single-stranded homologous DNA template. Here we review the current state of CRISPR/Cas-based gene editing and provide a practical guide to applying the CRISPR/Cas and rAAV system for highly efficient, time- and cost-effective gene targeting.
authors.affiliation: Department of Cellular and Molecular Medicine, University of California , San Diego, La Jolla, California.
authors.forename: Manuel
authors.initials: M
authors.lastname: Kaulich
authors.affiliation: Department of Cellular and Molecular Medicine, University of California , San Diego, La Jolla, California.
authors.forename: Steven F
authors.initials: SF
authors.lastname: Dowdy
mesh_terms: D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004247:DNA / Q000235:genetics; D000229:Dependovirus / Q000235:genetics*; D005818:Genetic Engineering; D005822:Genetic Vectors; D011995:Recombination, Genetic
chemical_list: D004247:DNA