title: Efficient conditional knockout targeting vector construction using co-selection BAC recombineering (CoSBR).
abstract: A simple and efficient strategy for Bacterial Artificial Chromosome (BAC) recombineering based on co-selection is described. We show that it is possible to efficiently modify two positions of a BAC simultaneously by co-transformation of a single-stranded DNA oligo and a double-stranded selection cassette. The use of co-selection BAC recombineering reduces the DNA manipulation needed to make a conditional knockout gene targeting vector to only two steps: a single round of BAC modification followed by a retrieval step.
authors.affiliation: Genentech, Inc., Department of Molecular Biology, 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Newman
authors.affiliation: Genentech, Inc., Department of Molecular Biology, 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Merone
authors.initials: M
authors.lastname: Roose-Girma
authors.affiliation: Genentech, Inc., Department of Molecular Biology, 1 DNA Way, South San Francisco, CA 94080, USA sorenw@gene.com.
authors.forename: Søren
authors.initials: S
authors.lastname: Warming
mesh_terms: D022202:Chromosomes, Artificial, Bacterial; D055786:Gene Knockout Techniques / Q000379:methods*; D005813:Genes, Synthetic; D005818:Genetic Engineering; D009841:Oligonucleotides; D011995:Recombination, Genetic
chemical_list: D009841:Oligonucleotides