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Find your way in pre-engineered cell lines!
Our cGPS® Targeted Integration (TI) kits contain an engineered cell line, which carry the recognition site of a natural occurring Meganuclease as a single copy. The Meganuclease is co-transfected with an Integration Matrix carrying your gene of interest (GOI). After recognizing a pre-integrated sequence in the cell genome (the nuclease 'landing pad'), the Meganuclease binds to the DNA and induces a DNA double-strand break. Natural repair mechanisms in the cell sense the damage and trigger homologous recombination to enable DNA repair. The co-transfected GOI, flanked by homology regions, is used as a template for the repair. The GOI is stably integrated at the Meganuclease recognition site.
Our cGPS® TI Kits comprise all necessary reagents for optimal targeted transgene integration in engineered cell lines:
- cGPS® Cell Line: the engineered cell line contains a “Landing Pad” (bearing a natural Meganuclease recognition site) stably integrated in a single copy, to support a highly efficient targeted integration two-step selection process;
- Natural Meganuclease: recognizes and cleaves a unique recognition site within the Landing Pad of an engineered cGPS® Cell Line;
- Integration Matrix: a plasmid that contains a Multiple Cloning Site (MCS) to clone your gene of interest downstream of a promoter. The transgene expression cassette is flanked by left and right homology arms to drive homologous recombination for targeted transgene integration at the Meganuclease recognition site.
Each individual TI kit component can be purchased separately .
cGPS® TI Kits generate more than 98% of targeted clones after the 2 step selection process. About 80% of the targeted clones are “pure” targeted clones, i.e. only contain a single transgene expression cassette at the targeted genomic site, without any additional randomly integrated copy.
cGPS® TI kits guarantee the efficient selection of stable and isogenic clones with highly stable expression within 3 weeks, even in the absence of selection agents.
Find within the pIM store a collection of Integration Matrices for a full-range of downstream applications, including gene expression regulation, gene tagging, integration of multiple expression cassettes at the same genomic locus, and many more.