Efficient conditional knockout targeting vector construction using co-selection BAC recombineering (CoSBR)

Robert J Newman; Merone Roose-Girma; Søren Warming

doi:10.1093/nar/gkv600

Efficient conditional knockout targeting vector construction using co-selection BAC recombineering (CoSBR)

Nucleic Acids Res. 2015 Oct 30;43(19):e124. doi: 10.1093/nar/gkv600. Epub 2015 Jun 18.

Authors

Robert J Newman¹, Merone Roose-Girma¹, Søren Warming²

Affiliations

¹ Genentech, Inc., Department of Molecular Biology, 1 DNA Way, South San Francisco, CA 94080, USA.
² Genentech, Inc., Department of Molecular Biology, 1 DNA Way, South San Francisco, CA 94080, USA [email protected].

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chromosomes, Artificial, Bacterial*
Gene Knockout Techniques / methods*
Genes, Synthetic
Genetic Engineering
Oligonucleotides
Recombination, Genetic*

Substances

Oligonucleotides