TARGATT™ Genome Editing

Recent advances in gene editing technologies have greatly increased our ability to make precise changes in the genome of cells or embryos and have eliminated major stumbling blocks in the generation of more sophisticated transgenic animal and cell line models. 

Applied StemCell’s proprietary, TARGATT™ allows for the site-specific integration of large DNA fragments, more efficiently and faster. The novel TARGATT™ technology uses the PhiC31 bacteriophage integrase to mediate an irreversible recombination between a pre-engineered “attP” docking-site and an attP recognition-sequence on the donor vector “attB”, for stable gene integration. The attP docking sites can be engineered into a specific or intergenic, transcriptionally active genomic locus (safe harbor locus), using CRISPR/Cas9. The transgene integration is site-specific, stable and with guaranteed expression.

TARGATT™ gene editing platform is very versatile and can be adapted to generate large fragment knock-in animal or cell line models from various species. This technology circumvents problems associated with random integration such as position effect, and gene silencing or instability due to integration of multiple copies of the transgene.

With Applied StemCell’s two complementary gene editing technologies, TARGATT™ and CRISPR/Cas9, we can generate a variety of advanced genetic modification in animals and cell lines to expand the scope of biomedical research and discoveries.

Advantages of TARGATT™ Technology:

• High integration efficiency (up to 40%)
• Large transgene knock-in (up to 22 kb)
• Reduced time and cost
• Guaranteed, high level expression of the transgene
• Site-specificity allows a precise comparison of the effects of the transgenes among different lines
• Site-specific knock-in at pre-selected locus overcomes challenges associates with random integration:
• Eliminates position effect
• Integration at intergenic region ensures that no internal genes are interrupted
• Single copy gene integration eliminates repeat-induced gene silencing and genomic instability

Applications:

• Transgene overexpression models (Ex. For bioproduction)
• Humanized animal models (large gene insertion)
• Reporter gene insertion models
• Inducible expression models (Ex. Tet-regulatory systems)
• Cre - driver lines