TARGATT™ mESC Master Cell Line & Knock-In Kit
Catalog # :
1 Kit (2 reactions)
The TARGATT™ Knock-in Mouse Cell Line Generation Kit is designed to create site-specific, knock-in mouse embryonic stem (ES) cells at a defined chromosomal locus in a fast, highly efficient way over traditional methods. The kit includes a mouse ES cell line of C57BL/6 strain background (AST-0020) with a TARGATT™ docking site (attP) at a transcriptionally active locus (the H11 locus on chromosome 11), and a plasmid for TARGATT™ integrase expression and a cloning vector to construct a donor DNA plasmid. With this method, large DNA fragments can be inserted at the H11 locus at high efficiency with robust transgene expression.
The TARGATT™ system takes advantage of the ΦC31 integrase enzyme from a Streptomyces phage. Integrases catalyze irreversible recombination between appropriate attB and attP sites. The ES cells included in the kit have been engineered with attP sites inserted at the H11 locus, previously characterized to confer high level transgene expression and is in an intergenic region. The DNA integration vector in the kit has the other ΦC31 integrase cognate site, attB. After transfecting the TARGATT™ ES cells with a vector expressing integrase and a DNA integration vector containing a gene of interest, recombination between attP site at the H11 locus and attB site on the integration vector results in transgene insertion at the H11 locus.
All reagents supplied here are sufficient for up to 2 rounds of transfection according to the protocol below. Reagents are also available separately from Applied StemCell.
We also provide custom TARGATT™ Master Cell Line Generation Service for “docking-site” site knock-in in your cell line and locus of choice as well as transgene knock-in into the TARGATT™ Master Cell Line.