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)

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Publisher's URL: http://dx.doi.org/10.1371/journal.pone.0019537


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
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 > Institute of Molecular Cell and Systems Biology
Journal Name:PLoS ONE
Publisher:Public Library of Science
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

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
378941Recombinase-mediated site-specific gene integration in mammalian somatic cellsWilliam StarkBiotechnology and Biological Sciences Research Council (BBSRC)BB/C003543/1Institute of Molecular Cell and Systems Biology