Zinc finger recombinases with adaptable DNA sequence specificity

Proudfoot, C., McPherson, A.L., Kolb, A.F. and Stark, W.M. (2011) Zinc finger recombinases with adaptable DNA sequence specificity. PLoS ONE, 6(4), e19537. (doi: 10.1371/journal.pone.0019537) (PMID:21559340) (PMCID:PMC3084882)

Abstract

Site-specific recombinases have become essential tools in genetics and molecular biology for the precise excision or integration of DNA sequences. However, their utility is currently limited to circumstances where the sites recognized by the recombinase enzyme have been introduced into the DNA being manipulated, or natural 'pseudosites' are already present. Many new applications would become feasible if recombinase activity could be targeted to chosen sequences in natural genomic DNA. Here we demonstrate efficient site-specific recombination at several sequences taken from a 1.9 kilobasepair locus of biotechnological interest (in the bovine beta-casein gene), mediated by zinc finger recombinases (ZFRs), chimaeric enzymes with linked zinc finger (DNA recognition) and recombinase (catalytic) domains. In the "Z-sites" tested here, 22 bp casein gene sequences are flanked by 9 bp motifs recognized by zinc finger domains. Asymmetric Z-sites were recombined by the concomitant action of two ZFRs with different zinc finger DNA-binding specificities, and could be recombined with a heterologous site in the presence of a third recombinase. Our results show that engineered ZFRs may be designed to promote site-specific recombination at many natural DNA sequences

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: McQue, Mrs Arlene and Proudfoot, Dr Christopher and Stark, Professor Marshall
• Authors: Proudfoot, C., McPherson, A.L., Kolb, A.F., and Stark, W.M.
• College/School: College of Medical Veterinary and Life Sciences > School of Molecular Biosciences
• Journal Name: PLoS ONE
• Publisher: Public Library of Science
• ISSN: 1932-6203
• ISSN (Online): 1932-6203
• Published Online: 29 April 2011
• Copyright Holders: Copyright © 2011 The Authors
• First Published: First published in PLoS ONE 6(4):e19537
• Publisher Policy: Reproduced in accordance with the copyright policy of the publisher

Project Code	Award No	Project Name	Principal Investigator	Funder's Name	Funder Ref	Lead Dept
37894	1	Recombinase-mediated site-specific gene integration in mammalian somatic cells	William Stark	Biotechnology and Biological Sciences Research Council (BBSRC)	BB/C003543/1	Institute of Molecular Cell and Systems Biology