phiC31 Integrase Vector System One-step Gene Addition Technology

NEW from System Biosciences (SBI) the phiC31 Integrase Cloning system for mammalian genome engineering applications. This system provides the researcher with the tools to perform non-viral, transfection-based experiments that result in
single-copy insertion of transgenes of interest with minimal risk of off-target effects, and can be easily adapted for in vivo applications. 


  • Single-copy, non-viral delivery system
  • Simple and effective way to generate stable cell lines and to genetically modify animals
  • Sustainable transgene expression (sometimes an issue for viral systems)
  • Low risk of insertional mutagenesis (which can often lead to oncogenic phenotype or chromosomal
  • abnormalities)
  • Ideal for stable transgene delivery to primary cells (which are often much more sensitive to random viral
  • integration effects)
  • Optimal for translating to in vivo applications (including gene therapy and regenerative medicine).
  • Single transfection protocol
  • Works in humans, mice, rats, pigs, chickens, cows, flies, frogs, and yeast (i.e. most mammalian and
  • eukaryotic model systems)
  • Extensive publication record for use in vitro and in vivo

Nonviral transgene delivery

One-step gene addition technology

The phiC31 integrase is a sequence-specific recombinase encoded within the genome of the bacteriophage phiC31. The phiC31 integrase mediates recombination between two 34 base pair sequences termed attachment sites (att), one found in the phage and the other in the bacterial host. This serine integrase has been show to function efficiently in many different cell types including mammalian cells. In the presence of phiC31 integrase, an attB- containing donor plasmid can be unidirectional integrated into a target genome through recombination at sites with sequence similarity to the native attP site (termed pseudo-attP sites). phiC31 integrase can integrate a plasmid of any size, as a single copy, and requires no cofactors. The integrated transgenes are stably expressed and heritable.


  • Nonviral transgenesis
  • Single-copy integration
  • Preferred integration at active transcription sites
  • Unlimited cargo capacity
  • Easily make stable cell lines using transfection-based methods
  • Suitable for engineering Stem Cells and In vivo animal models
Product Size Catalogue No.
PhiC31 Integrase Expression Plasmid 10ug FC200PA-1
pFC-MCS-pA-SV40-Neo PhiC31 Donor Vector 10 ug FC500A-1
pFC-CMV-MCS-pA-SV40-Neo PhiC31 Donor Vector 10 ug FC501A-1
pFC-CMV-GFP-SV40-Neo Positive Control Donor Vector 10 ug FC520A-1
pFC-EF1-MCS-pA-PGK-RFP-T2A-Puro PhiC31 Donor Vector 10 ug FC550A-1
pFC-PGK-MCS-pA-EF1-GFP-T2A-Puro PhiC31 Donor Vector 10 ug FC551A-1


Online Resources

  • phiC31 Integrase Vector System User Manual
  • WEBINAR Slide set for phiC31 Integrase Vector System

Technical References

  • Karow M, Chavez CL, Farruggio AP, Geisinger JM, Keravala A, Jung WE, Lan F, Wu JC, Chen-Tsai Y, Calos MP. Site-specific recombinase strategy to create induced pluripotent stem cells efficiently with plasmid DNA. Stem Cells 29(11):1696-704.
  • Chalberg TW, Portlock JL, Olivares EC, Thyagarajan B, Kirby PJ, Hillman RT, Hoelters J, Calos MP. Integration specificity of phage phiC31 integrase in the human genome. J Mol Biol. 2006 Mar 17;357(1):28-48.
  • Lan F, Liu J, Narsinh KH, Hu S, Han L, Lee AS, Karow M, Nguyen PK, Nag D, Calos MP, Robbins RC, Wu JC. Safe genetic modification of cardiac stem cells using a site-specific integration technique. Circulation. 2012 Sep 11;126(11 Suppl 1):S20-8.
  • Hillman RT, Calos MP. Site-specific integration with bacteriophage phiC31 integrase. Cold Spring Harb Protoc. 2012 May 1;2012(5).
  • Chavez CL, Keravala A, Chu JN, Farruggio AP, Cuellar VE, Voorberg J, Calos MP. Long-term expression of human coagulation factor VIII in a tolerant mouse model using the phiC31 integrase system. Hum Gene Ther. 2012 Apr;23(4):390-8.
  • Woodard LE, Keravala A, Jung WE, Wapinski OL, Yang Q, Felsher DW, Calos MP. Impact of hydrodynamic injection and phiC31 integrase on tumor latency in a mouse model of MYC-induced hepatocellular carcinoma. PLoS One. 2010 Jun 29;5(6):e11367.


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