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phiC31 Integrase Vector System: One-Step Gene Addition Technology

Nonviral transgene delivery
• Single-copy integration minimizing off-target effects and insertional mutations
• Preferred integration at active transcription sites
• Sustainable transgene expression
• Unlimited cargo capacity
• Easily make stable cell lines using transfection-based methods
• Ideal for stable transgene delivery to primary cells
• Suitable for engineering stem cells and in vivo animal models enabling in vivo applications incl. gene therapy and regenerative medicine approaches
• Works in human cells and most mammalian and eukaryotic model systems (e.g. mouse, rat, pig, chicken, cow, flies, frogs, and yeast)
• Extensive publication record for in vitro and in vivo use


One-step gene addition technology

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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.



Choice of phiC31 Integrase Vectors

phiC31 integrase expression construct
All of the phiC31 donor vectors require co-transfection with the phiC31 integrase expression vector to catalyze the donor vector integration.

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This construct (Cat#FC200A-1-SBI) has a codon-optimized phiC31 cDNA expressed by CMV for transient expression of the integrase to inert the donor expression vectors into pseudo attP genomic sites using the attB sequences in the donor vectors. This construct has a kanamycin marker for propagation in bacteria.



Dual-direction promoter donor vectors
The dual promoter phiC31 vectors are available in two formats: The pFC-EF1-MCS- PGK-RFP-Puro vector (cat# FC550A-1-SBI) has EF1a driving your gene´s expression cloned into the MCS and PGK expressing the RFP-Puro combo markers.
The alternative format pFC-PGK-MCS-EF1-GFP-Puro (cat#FC551A-1-SBI) utilizes PGK to express your gene cloned into the MCS and has EF1a driving the GFP-Puro combo markers. Both PGK and EF1a promoters are well-validated to have a broad range of activity across many cell types, including stem cells.

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Single promoter and promoterless vector options

The pFC-CMV-MCS-SV40-Kan/Neo phiC31 vector (cat# FC501A-1-SBI) features CMV driving the expression of your gene cloned into the MCS.

pFC-CMV-GFP-SV40-Kan/Neo phiC31 (cat# FC520A-1-SBI) vector features CMV driving the expression of GFP as a positive control construct.

The pFC-MCS-SV40-Kan/Neo phiC31 integrase vector (cat# FC500A-1-SBI) is an entry-level empty vector to allow any promoter and gene cassette to be inserted at the MCS site.

All 3 vectors have a downstream SV40 promoter expressing the combo Kan/Neo marker for propagation in bacteria with kanamycin and also neomycin selection in mammalian cells.

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Related Links

Literature References
Experimental data showing phiC31 based integration
phiC31 based IPSC generation


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Brochure about phiC31 Integrase Vector System

 

 

Description Cat# Size Price    
PhiC31 Integrase Expression Plasmid FC200PA-1-SBI 10 ug 566 € DETAILS   Add to Cart 
pFC-EF1-MCS-pA-PGK-RFP-T2A-Puro PhiC31 Donor Vector (dual promoter) FC550A-1-SBI 10 ug 758 € DETAILS   Add to Cart 
pFC-PGK-MCS-pA-EF1-GFP-T2A-Puro PhiC31 Donor Vector (dual promoter) FC551A-1-SBI 10 ug 758 € DETAILS   Add to Cart 
pFC-CMV-MCS-pA-SV40-Neo PhiC31 Donor Vector (single promoter, no fluorescent protein marker) FC501A-1-SBI 10 ug 758 € DETAILS   Add to Cart 
pFC-CMV-GFP-SV40-Neo Positive Control Donor Vector FC520A-1-SBI 10 ug 758 € DETAILS   Add to Cart 
pFC-MCS-pA-SV40-Neo PhiC31 Donor Vector (promoterless) FC500A-1-SBI 10 ug 758 € DETAILS   Add to Cart 
pFC-CAG-MCS-WPRE-pA-SV40-Neo PhiC31 Donor Vector (single promoter, no fluorescent protein marker) FC600A-1-SBI 10 ug 758 € DETAILS   Add to Cart 

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