Safe Harbor Locus Master iPSC Genome Editing Service

Custom “Master” iPSC Cell Line Generation using TARGATT™ technology! The TARGATT™ Master Human iPSC Line contains an attP “docking site” at the hROSA26 safe harbor genomic locus. Any gene of interest can be stably inserted at the docking site using a corresponding “attB” containing TARGATT™ donor plasmid, with guaranteed gene expression. The efficiency of this phiC31 integrase-mediated recombination is up to 100% efficiency with drug selection and up to 30% without drug selection.

Our cell line model generation experts can also engineer your safe harbor locus knock-in iPSC lines using CRISPR/Cas9 for a multi-technology approach to generate a cell line model for your specific research needs.

Products and Services
Application Notes

ASC engineered Site-Specific Knock-in of TARGATT™ sites, with mCherry reporter (left). Original Cell Line: hiPSC (right).

mCherry reporter - iPSC Generation Original Cell Line: hiPSC - iPSC Generation
Technical Details

Safe harbor locus master cell lines are ideal for applications using reporter lines for cell tracking, for cell purification using surface markers, and for directed differentiation of iPSCs using cell-specific promoters.

iPSC lines with cell-specific promoters:

  • Express cell-type specific transcription factors to induce differentiation of specific cell lineages

  • Reporters to track cell differentiation process

Potential Projects:

  • iPSC lines with excisable Cas9 or nickase for efficient CRISPR gene modification

  • High-efficiency, high purity cells differentiated from iPSC lines

Applied StemCell can also create Site-Specific Knock-in cell lines for primary cells or other cell lines.

  • Karow, M., et al. (2011). Stem Cells, 29(11), 1696–1704.

  • Zhu, F., et al. (2014). Nucleic acids research, 42(5), e34-e3

  • Lizarraga, S. B., Maguire, A. M., Ma, L., van Dyck, L. I., Wu, Q., Nagda, D., ... & Cowen, M. H. (2018). Human neurons from Christianson syndrome iPSCs reveal allele-specific responses to rescue strategies. bioRxiv, 444232.
  • Tanaka, H., Kondo, K., Chen, X., Homma, H., Tagawa, K., Kerever, A., ... & Fujita, K. (2018). The intellectual disability gene PQBP1 rescues Alzheimer’s disease pathology. Molecular Psychiatry, 1.4
  • Selvan N., George, S., Serajee, F. J., Shaw, M., Hobson, L., Kalscheuer, V. M., ... & Schwartz, C. E. (2018). O-GlcNAc transferase missense mutations linked to X-linked intellectual disability deregulate genes involved in cell fate determination and signaling. Journal of Biological Chemistry, jbc-RA118.

  • Chai, S., Wan, X., Ramirez-Navarro, A., Tesar, P. J., Kaufman, E. S., Ficker, E., ... & Deschênes, I. (2018). Physiological genomics identifies genetic modifiers of long QT syndrome type 2 severity. The Journal of clinical investigation128(3).

  • Seigel, G. M., et al. (2014). Comparative Analysis of ABCG2+ Stem-Like Retinoblastoma Cells and Induced Pluripotent Stem Cells as Three-Dimensional Aggregates. Investigative Ophthalmology & Visual Science55(13), 3068-3068.

  • Comley, J. (2016). CRISPR/Cas9 - transforming gene editing in drug discovery labs. Drug Discovery Weekly. Fall 2016; 33-48.

Have Questions?

An Applied StemCell technical expert is happy to help, contact us today!