A genetic screen identifies novel non-compatible loxP sites
- 15 July 2002
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 30 (14) , 3067-3077
- https://doi.org/10.1093/nar/gkf421
Abstract
The ability of the Cre/lox system to make precise genomic modifications is a tremendous accomplishment. However, recombination between cis-linked heterospecific lox sites limits the use of Cre- mediated exchange of DNA to systems where genetic selection can be applied. To circumvent this problem we carried out a genetic screen designed to identify novel mutant spacer-containing lox sites displaying enhanced incompatibility with the canonical loxP site. One of the mutant sites recovered appears to be completely stable in HEK293 cells constitutively expressing Cre recombinase and supports recombinase-mediated cassette exchange (RMCE) in bacteria and mammalian cell culture. By preventing undesirable recombination, these novel lox sites could improve the efficiency of in vivo gene transfer.Keywords
This publication has 25 references indexed in Scilit:
- Selection-Marker-Free Modification of the Murine β-Casein Gene Using a lox2722 SiteAnalytical Biochemistry, 2001
- Site-specific chromosomal integration in mammalian cells: highly efficient CRE recombinase-mediated cassette exchangeJournal of Molecular Biology, 1999
- Cre-stimulated recombination atloxP-containing DNA sequences placed into the mammalian genomeNucleic Acids Research, 1989
- THE MECHANISM OF CONSERVATIVE SITE-SPECIFIC RECOMBINATIONAnnual Review of Genetics, 1988
- Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1.Proceedings of the National Academy of Sciences, 1988
- A human beta-actin expression vector system directs high-level accumulation of antisense transcripts.Proceedings of the National Academy of Sciences, 1987
- Functional expression of the cre-lox site-specific recombination system in the yeast Saccharomyces cerevisiae.Molecular and Cellular Biology, 1987
- The role of theloxPspacer region in PI site-specific recombinationNucleic Acids Research, 1986
- P1 site-specific recombination: nucleotide sequence of the recombining sites.Proceedings of the National Academy of Sciences, 1982
- A novel role for site-specific recombination in maintenance of bacterial repliconsCell, 1981