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The TARGATT™ HEK293 Master Cell Line and Knockin Kit was designed for fast and site-specific knockin in HEK293 cells, using an easy-to-use gene knockin 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 knockin and uniform, stable gene expression.
The TARGATT™ HEK293 master cell line and knockin 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*.
Advantages of using TARGATT™ Master Cell Lines for gene knockin: