The TARGATT™-HEK293 Master Cell Line and Knock-in Kit was designed for fast and site-specific knock-in in HEK293 cells, using an easy-to-use gene knock-in approach. The master cell line provided in this kit contains an “attP” integrase-recognition landing pad engineered into the hH11 safe harbor locus in the genome. The kit also contains a cloning plasmid containing a corresponding “attB” sequence into which any gene of interest can be cloned (under control of the CMV promoter or a promoter-of-choice). The expression of the integrase (provided as an integrase plasmid) mediates the stable integration of the transgene into the master cell line (Figure 1). The TARGATT™ integrase technology enables highly efficient (>40 % without enrichment and ~90 % with selection), and site-specific DNA integration without disruption of internal genes. The TARGATT™-HEK293 master cell line can therefore be used for building mammalian cell libraries with site-specific, single transgene gene knock-in and uniform, stable gene expression.
Advantages of using TARGATT™ Master Cell Lines for gene knock-in:
– High efficiency Integration
– Site-specific, stable knock-in cell line generation
– Single copy gene integration into safe harbor locus
– Gene expression from an active, intergenic locus
– Low off-target integration
The TARGATT™-HEK293 master cell line and knock-in kit are suitable for research applications involving directed-evolution of proteins (vaccine development, drug screening, cell-based gene therapy), genome-wide screening, and other stable cell line generation applications. If you are interested to get your own MASTER cell line (including stem cells), please contact us to discuss your project.