FAK CRISPR Activation Plasmid (m): sc-420280-ACT

Protein tyrosine kinase 2 (Ptk2) encodes focal adhesion kinase (FAK), a non-receptor tyrosine kinase that integrates signals from integrins and growth factor receptors to coordinate focal adhesion turnover and actin cytoskeleton remodeling. In mouse cells, FAK regulates cell adhesion, migration, polarity, and survival through pathways including PI3K–AKT, MAPK/ERK, Rho-family GTPase signaling, and SRC-dependent phosphorylation cascades. FAK also interfaces with mechanotransduction programs that couple extracellular matrix stiffness to transcriptional responses and cellular contractility. Dysregulated FAK signaling is associated with altered tissue remodeling, inflammatory microenvironments, and invasive phenotypes, making Ptk2 a widely studied node in models of tumor progression, fibrosis, and vascular biology.

FAK CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ptk2 expression without altering the underlying DNA sequence.

FAK CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ptk2 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 Ptk2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FAK expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ptk2 locus and enabling the study of FAK-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FAK pathway restoration in tumor cells with silenced or reduced Ptk2 expression.

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