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Open Access Research

Phage integrases for the construction and manipulation of transgenic mammals

Roger P Hollis126, Stephanie M Stoll136, Christopher R Sclimenti14, Jennifer Lin5, Yanru Chen-Tsai5 and Michele P Calos1*

Author Affiliations

1 Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305-5120, USA

2 Present address: Children's Hospital of Los Angeles, Research Immunology/BMT, Mail Stop #62, 4650 Sunset Boulevard, Los Angeles, CA 90027, USA

3 Present address: University of California San Francisco, Department of Surgery, 513 Parnassus Ave, HSW 1622, Box 0522, San Francisco, CA 94122-0522, USA

4 Present address: Poetic Genetics LLC, 863B Mitten Rd., Burlingame, CA 94010, USA

5 Stanford Transgenic Research Facility, Stanford University School of Medicine, Stanford, CA 94305-5120, USA

6 Joint first authors

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Reproductive Biology and Endocrinology 2003, 1:79  doi:10.1186/1477-7827-1-79

Published: 7 November 2003

Abstract

Phage integrases catalyze site-specific, unidirectional recombination between two short att recognition sites. Recombination results in integration when the att sites are present on two different DNA molecules and deletion or inversion when the att sites are on the same molecule. Here we demonstrate the ability of the φC31 integrase to integrate DNA into endogenous sequences in the mouse genome following microinjection of donor plasmid and integrase mRNA into mouse single-cell embryos. Transgenic early embryos and a mid-gestation mouse are reported. We also demonstrate the ability of the φC31, R4, and TP901-1 phage integrases to recombine two introduced att sites on the same chromosome in human cells, resulting in deletion of the intervening material. We compare the frequencies of mammalian chromosomal deletion catalyzed by these three integrases in different chromosomal locations. The results reviewed here introduce these bacteriophage integrases as tools for site-specific modification of the genome for the creation and manipulation of transgenic mammals.