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TARGATT™ CHO-K1 Knock-in Kit (A2)

SKU AST-1405 Categories , ,

TARGATT™ CHO-K1 Knock-in Kit (A2)

Insert confidently with our TARGATT™ Knock-in Kit:

  • High efficiency, unidirectional integration
  • Site-specific, stable knock-in cell line generation
  • Single-copy gene integration into a safe harbor locus
  • Gene expression from an active, intergenic locus
  • Easy-to-use protocol requiring transfection of only one (donor) plasmid

Overview

The TARGATT™ CHO-K1 Knock-in Kit (A2) is designed for fast and site-specific knock-in in Chinese Hamster Ovary (CHO) cells using an easy-to-use gene knock-in approach. The master cell line provided in this kit contains both the “attP” docking site and the PhiC31 integrase expression cassette engineered into the CHO ASC2 (A2) safe harbor locus in the genome. Any gene of interest can be cloned into the provided TARGATT™ “attB” cloning plasmid (under control of the strong CAG promoter or promoter-of-choice) 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.

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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 master cell line. The TARGATT™CHO-K1 (A2) Master Cell Line was engineered with the attP landing pad at the proprietary ASC2 (A2) safe harbor locus. The TARGATT™ plasmid containing the integrase recognition site, attB is used to clone the transgene. The integrase catalyzes an irreversible reaction between the attP site in the genome and attB site in the donor vector, resulting in sitespecific integration of the gene of interest at the selected locus. The cells containing the gene of interest can be enriched using ganciclovir negative selection.

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