Brk CRISPR Activation Plasmid (h): sc-401127-ACT

Protein tyrosine kinase 6 (PTK6), also known as Brk, is a non-receptor tyrosine kinase implicated in regulation of epithelial signaling, cytoskeletal dynamics, and cell migration. Brk modulates growth factor and cytokine-driven pathways, intersecting with EGFR/HER family signaling and downstream MAPK and PI3K/AKT network components to influence proliferation and survival programs. PTK6 activity and localization have been linked to altered phosphorylation of adaptor proteins and transcriptional regulators, shaping cellular responses to stress and differentiation cues. Dysregulated PTK6 expression has been reported across multiple tumor contexts, making it a useful molecular node for mechanistic studies of oncogenic signaling, invasion phenotypes, and pathway rewiring.

Brk CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTK6 expression without altering the underlying DNA sequence.

Brk CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTK6 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 PTK6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Brk expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTK6 locus and enabling the study of Brk-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Brk pathway restoration in tumor cells with silenced or reduced PTK6 expression.

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