The TARGATT™ hiPSC Master Cell Line and transgenic kit were designed for fast and site-specific knock-in in human iPSC 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 CAG promoter). 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 site-specific integration after antibiotic enrichment, without disruption of internal genes. The TARGATT™ iPSC master cell line and knockin kit are ideal for single transgene knockin and uniform, stable expression of your protein*.
The TARGATT™ iPSC Master Cell Line has been engineered from our well-characterized, karyotype normal, control iPSC line, ASE-9211. The ASE-9211 parental iPSC line was reprogrammed using episomal factors from fibroblasts obtained with full consent from a neonatal, African-American male donor. Both the parental ASE-9211 line and the TARGATT™ iPSC Master Cell Line have been characterized for pluripotency biomarkers, normal karyotype, and directed-differentiation to three germ layers as a validation of functional pluripotency.
The TARGATT™ iPSC master cell line and knockin kit are suitable for research applications involving site-specific large transgene knockin, gene overexpression, and other stable cell line generation applications.
H11 Safe-Harbor Locus
The H11 safe-harbor locus is in an intergenic region between two highly expressed genes. ASC has identified orthologous sequences of H11 in various cell lines (e.g., human cells, HEK293 cells, iPS cells, and CHO cells) and animals (e.g., mice, pigs, and rats). Insertion at the H11 site leads to little or no phenotypic change. Also, integration does not affect any endogenous gene expression or function, and medium to high levels of transgene expression have been observed.