TARGATT™ Cell Line Model Generation
Applied StemCell Inc.’s proprietary TARGATT™ technology enables highly efficient site-specific gene integration in mammalian cells. This technology uses ФC31 integrase to insert any gene of interest into a docking site, pre-engineered in an intergenic and transcriptionally active genomic locus.
Our TARGATT™ technology improves several aspects in the generation of transgenic cell lines:
(1) High integration efficiency mediated by ФC31 integrase reduces time and cost
(2) Site-specific integration at a pre-selected genomic locus eliminates position effect and ensures high level expression of the transgene
(3) Integration at intergenic region ensures that no internal genes are interrupted
(4) Single copy gene integration eliminates repeat-induced gene silencing and genomic instability
(5) Site-specific integration allows a precise comparison of the effects of the transgenes among different lines.
The TARGATT™ technology can be utilized for a variety of applications including reporter gene expression, gene knockdown, conditional expression, disease cell and animal models.
Figure 1. Scheme for site-specific gene insertion knock-in using TARGATTTM Knock-in Mouse Cell Line Generation Kit. The TARGATTTM ES cells contain three tandem attP “docking” sites knocked into H11 locus. A mix of the TARGATTTM integrase vector and integration vector containing the gene of interest (e.g., GFP) and an attB site was electroporated into the TARGATTTM ES cells. TARGATTTM integrase catalyzes recombination between attP and attB sites, resulting in integration of GFP.
Description of the technology
- Zhu, F., et al. (2014). Nucleic acids research, 42(5), e34-e34.
- Tasic, B., et al. (2011). PNAS USA, 108(19), 7902–7907.
Commentary, comparison with other transgenic methods
- Rossant, J., et al. (2011). PNAS USA 108(19), 7659–7660.
Tet inducible mice generated by TARGATT™
- Fan, X., et al. (2012). Endocrinology, 153(11), 5637–5644.
Advantage of Hipp11 (H11) locus
- Hippenmeyer, S., et al. (2010). Neuron, 68(4), 695–709.
Applications for mice generated by TARGATT™
- Booze, M. L., et al. (2016). Free Radical Biology and Medicine, 99: 533-543.
- Feng, D., et al. (2016). The Journal of Clinical Investigation, 126(6).
- Park, K. E., et al. (2016). International journal of molecular sciences,17(6), 810.
- Sun, N., et al. (2015). Measuring in vivo mitophagy. Molecular cell, 60(4), 685-696
- Guenther, CA., et al. (2014). Nature genetics, 46(7), 748-752
- Devine, WP., et al. (2014). eLife, 3, e03848.
- Villamizar, C. A. (2014) UT GSBS Dissertations and These (Open Access). Paper 508 (2014)
- Fogg, PC., et al. (2014). Journal of molecular biology, 426(15), 2703-2716
- Chen-Tsai, RY. (2014) Chinese Science. 59:1-6.