TARGATT™ HEK293 Master Cell Line & Knock-in Kit

SKU :
AST-1301
Catalog # :
AST-1301
Size :
1 Kit
$1,500.00
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Overview

* Price listed is for academic customers. Industry clients, please inquire

The TARGATT™-HEK293 Master Cell Line and transgenic kit was designed for fast and site-specific knock-in in HEK293 cells, using an easy-to-use gene knock-in approach. The master cell line provided in this kit contains the “attP” docking-site and the PhiC31 integrase expression cassette engineered into the hROSA26 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 guarantees efficient DNA integration and high-level gene expression without disrupting internal genes. The TARGATT™-HEK293 master 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™-HEK293 master cell line and transgenic kit are suitable for research applications involving gene overexpression and high-level expression of recombinant proteins and other biologics in a rapidly expanding bioproduction industry and other applications*.

*TARGATT™ master cell lines can be generated in any cell line including stem cells. Please contact Applied StemCell for TARGATT™ cell line engineering services to generate a master cell line in a specific cell line of choice.

This product is for research purposes only.


TARGATT™ Master Cell Line

  • Chi, X., Zheng, Q., Jiang, R., Chen-Tsai, R. Y., & Kong, L. J. (2019). A system for site-specific integration of transgenes in mammalian cells. PLOS ONE14(7), e0219842.

Advantages of using TARGATT™ Master Cell Lines for gene knock-in:

  • Stable cell line generation
  • Single copy, site-specific gene integration into the safe harbor locus
  • Guaranteed gene expression from an active, intergenic locus
  • Footprint-free integration of gene of interest
  • Use of selection marker is not necessary but optional
  • Easy-to-use protocol requiring transfection of only one (donor) plasmid

Contents:

All cell lines and reagents provided in this kit are sufficient for transfection according to the given protocol:

  • TARGATT™-HEK293 Master Cell line (AST-1301-1)
  • TARGATT™ 22 (CAG-MCS-TK, RV-attB) cloning plasmid (AST-3062)
  • TARGATT™ 23 (CAG-GFP-TK, RV-attB) positive control plasmid (AST-3063)
  • Ganciclovir (AST-1300-2)

Schematic Representation of TARGATT™ Knock-in Strategy:

landingpage-ast-3101-hek293-schematic

Figure 1. Schematic representation of site-specific gene insertion using TARGATT™ Technology. The TARGATT™-HEK  Master Cell Line was engineered with the attP “docking” site at the hROSA26 safe harbor locus. The TARGATT™ integrase catalyzes an irreversible reaction between attP sites on genome and attB sites on donor vector, resulting in integration of gene of interest at the preselected locus.

GFP Expression 21 days Post-Transfection


landingpage-targatt-hek293-facs

Figure 2. The CAG-GFP vector was used to verify the fast knock-in the TARGATT™-HEK293 master cell line. An enriched GFP signal was shown under fluorescence microscopy. (Left) Bright field microscopy. (Middle) Immunofluorescence: GFP channel; 5X magnification. (Right) FACS analysis for GFP signal of cell population. A. Before GCV selection; 13.9%. B. After GCV selection; 89.7%. C. Control parental HEK293 line; 1.06%.