TARGATT™ Genome Editing

 

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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™ that 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.

SCHEMATIC-TARGATT-TECHNOLOGY

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 (Example: Tet-regulatory systems)
  • Cre - driver animal models - We can generate Cre animal models using promoters listed below or ANY promoter provided by customers (with available sequence) using our versatile TARGATT™ technology:

Choose the right gene editing technology

Applied StemCell uses two complementary gene editing technologies, CRISPR/Cas9 and our propriety site-specific integrase-based TARGATT™ technology for generating advanced cell line and animal models very efficiently and effectively.

Project Purpose

CRISPR/Cas9

TARGATT™

Knock-Out (KO)

Yes

 

Point Mutation

Yes

 

Conditional KO

Yes

 

Knock-In

(<200 Nucleotide ssODN Donor)

Yes

 

Knock-In Transgenes in

Safe Harbor Loci (>2kb)

Challenging

(but limitations on size)

Yes

 (up to 20kb)

Knock-In

 (Plasmid DNA)

Challenging

(but limitations on size)

Yes

 (2 steps: KI docking site; KI transgene)