Cortactin CRISPR Activation Plasmid (h): sc-400761-ACT

Human CTTN encodes cortactin, a Src substrate and F-actin–binding scaffold that promotes Arp2/3-dependent branched actin assembly and stabilizes dynamic actin networks at the cell cortex. Cortactin coordinates signaling from receptor tyrosine kinases and Src-family kinases with cytoskeletal remodeling through interactions with N-WASP, cofilin, and endocytic adaptors, influencing lamellipodia formation, focal adhesion turnover, and vesicle trafficking. Through these processes, CTTN is tightly linked to cell migration, invasion-like behavior, and mechanotransduction, and its dysregulation has been associated with altered motility programs and tumor progression phenotypes. As a result, CTTN is frequently studied in pathways governing actin dynamics, endocytosis, and extracellular matrix interactions.

Cortactin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CTTN expression without altering the underlying DNA sequence.

Cortactin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CTTN locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the CTTN transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cortactin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CTTN locus and enabling the study of Cortactin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cortactin pathway restoration in tumor cells with silenced or reduced CTTN expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.