Transgene excision in zebrafish using the phiC31 integrase
Article first published online: 21 JAN 2010
Copyright © 2010 Wiley-Liss, Inc.
Volume 48, Issue 2, pages 137–143, February 2010
How to Cite
Lister, J. A. (2010), Transgene excision in zebrafish using the phiC31 integrase. Genesis, 48: 137–143. doi: 10.1002/dvg.20595
- Issue published online: 3 FEB 2010
- Article first published online: 21 JAN 2010
- Manuscript Accepted: 15 NOV 2009
- Manuscript Revised: 13 NOV 2009
- Manuscript Received: 17 MAR 2009
- National Institute of Child Health and Human Development. Grant Number: R21 HD055311
- NIH-NCRR. Grant Number: P40 RR012546
Genome engineering strategies employing site-specific recombinases (SSRs) have become invaluable to the study of gene function in model organisms. One such SSR, the integrase encoded by the Streptomyces bacteriophage phiC31, promotes recombination between heterotypic attP and attB sites. In the present study I have examined the feasibility of the use of phiC31 integrase for intramolecular recombination strategies in zebrafish embryos. I report here that (1) phiC31 integrase is functional in zebrafish cells, (2) phiC31 integrase can excise a transgene cassette flanked by an attB and an attP site, analogous to a common use of the Cre/lox SSR system, (3) phiC31 integrase functions in the zebrafish germline, and (4) a phiC31 integrase-estrogen receptor hormone-binding domain variant fusion protein catalyzes attB-attP recombination in zebrafish embryos in a 4-hydroxytamoxifen-dependent manner, albeit less efficiently than phiC31 alone. These features should make this a useful approach for genome manipulations in the zebrafish. genesis 48:137–143, 2010. © 2010 Wiley-Liss, Inc.