Phage integrases for the construction and manipulation of transgenic mammalsReport as inadecuate




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Reproductive Biology and Endocrinology

, 1:79

First Online: 07 November 2003Received: 15 July 2003Accepted: 07 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.

Electronic supplementary materialThe online version of this article doi:10.1186-1477-7827-1-79 contains supplementary material, which is available to authorized users.

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Author: Roger P Hollis - Stephanie M Stoll - Christopher R Sclimenti - Jennifer Lin - Yanru Chen-Tsai - Michele P Calos

Source: https://link.springer.com/







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