Site-directed insertion of transgenes

Volume 23 of the series Topics in Current Genetics pp 211-239


Site-Specific Recombination Using PhiC31 Integrase

  • Jonathan M. GeisingerAffiliated withDepartment of Genetics, Stanford University School of Medicine
  • , Michele P. CalosAffiliated withDepartment of Genetics, Stanford University School of Medicine Email author 

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The integrase from phage φC31 of Streptomyces bacteria is an attractive recombinase for use in generating transgenic organisms and developing gene and cell therapeutic strategies. In nature, φC31 integrase mediates integration by interacting with specific sites in the phage and bacterial genomes. When applied to eukaryotes, φC31 integrase provides efficient unidirectional recombination between its own attB and attP sites or between an attB site on an incoming plasmid and a native genomic pseudo attP site that resembles attP. To date, the φC31 system has been used to generate stable transgenic organisms from multiple species, including plants, insects, and vertebrates. The features of the φC31 system also make it particularly amenable to therapeutic strategies. φC31 integrase has been used in potential therapies for numerous genetic diseases including hemophila, muscular dystrophy, and skin disorders. Additionally, the φC31 system has recently been used to modify human embryonic stem cells and to generate induced pluripotent stem cells. The φC31 system can also be combined with other recombinases to create advanced genome engineering strategies. In the future, the use of φC31 integrase may facilitate the development of new gene and cell therapies, as well as the generation of targeted transgenic organisms.


Gene therapy Induced pluripotent stem cells Phage integrase Recombinases Site-specific integration Transgenic organisms