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TARGATT™ CHO-K1 Knock-in Kit (H11)

SKU AST-1420 Categories , ,

TARGATT™ CHO-K1 Knock-in Kit (H11)

Advantages of using TARGATT™ master cell lines for gene knock-in:

  • High efficiency, unidirectional integration
  • Site-specific, stable knock-in cell line generation
  • Single copy gene integration into safe harbor locus
  • Gene expression from an active, intergenic locus
  • Easy-to-use protocol

Overview

The TARGATT™ CHO-K1 Knock-in Kit (H11) is designed for fast and site-specific knock-in in Chinese Hamster Ovary (CHO) cells with an easy-to-use approach. The master cell line provided in this kit contains both the “attP” docking site and the integrase expression cassette engineered into the CHO H11 safe harbor locus in the genome. Any gene of interest can be cloned into the provided TARGATT™ “attB” cloning plasmid and transfected into the master cell line for generating a stable knock-in cell line. The TARGATT™ integrase-based technology enables efficient DNA integration and high-level gene expression without disrupting internal genes. The TARGATT™ CHO-K1 cell line can therefore be used for uniform, site-specific gene knock-in, generation of isogenic cell lines, and amplification strategies for isolating high-expression cells, without the need for single-cell cloning.

The TARGATT™ CHO-K1 Knock-in Kit (H11) is suitable for research applications involving gene overexpression and high-level expression of recombinant proteins and other biologics in a rapidly expanding bioproduction industry, and for other applications.

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Our knock-in strategy

Figure 1. Schematic representation of TARGATT™ site-specific transgene integration mediated by integrase in TARGATT™ CHO-K1 (H11) master cell line. The TARGATT™ CHO-K1 (H11) master cell line was engineered with the attP landing pad at the Hipp11 (H11) safe harbor locus. The TARGATT™ plasmid containing the integrase recognition site, attB is used to clone the transgenes. The integrase catalyzes an irreversible reaction between the attP site in the genome and attB site in the donor vector, resulting in site-specific integration of the gene of interest at the selected locus. The cells that have stably knocked-in the donor plasmid can be enriched with hygromycin. Parental cells, random integrants and other undesired cells can be further eliminated via ganciclovir selection.

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