Site-Specific Genomic Integration in Mammalian Cells Mediated by Phage {phi}C31 Integrase

doi:10.1128/MCB.21.12.3926-3934.2001

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Molecular and Cellular Biology, June 2001, p. 3926-3934, Vol. 21, No. 12
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.12.3926-3934.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Site-Specific Genomic Integration in Mammalian Cells Mediated by Phage $phi$ C31 Integrase

Bhaskar Thyagarajan, Eric C. Olivares, Roger P. Hollis, Daniel S. Ginsburg, and Michele P. Calos^*

Department of Genetics, Stanford University School of Medicine, Stanford, California 94305-5120

Received 12 January 2001/Returned for modification 21 February 2001/Accepted 23 March 2001

We previously established that the phage $phi$ C31 integrase, a site-specific recombinase, mediates efficient integration in thehuman cell environment at attB and attP phage attachment siteson extrachromosomal vectors. We show here that phage attP sitesinserted at various locations in human and mouse chromosomes serveas efficient targets for precise site-specific integration. Moreover,we characterize native "pseudo" attP sites in the human and mousegenomes that also mediate efficient integrase-mediated integration.These sites have partial sequence identity to attP. Such sitesform naturally occurring targets for integration. This phage integrase-mediatedreaction represents an effective site-specific integration systemfor higher cells and may be of value in gene therapy and otherchromosome engineeringstrategies.

^* Corresponding author. Mailing address: Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305-5120.Phone: (650) 723-5558. Fax: (650) 725-1534. E-mail: calos{at}stanford.edu.

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Site-Specific Genomic Integration in Mammalian Cells Mediated by Phage C31 Integrase

Site-Specific Genomic Integration in Mammalian Cells Mediated by Phage $phi$ C31 Integrase