PYK2 CRISPR Activation Plasmid (h): sc-400708-ACT

PTK2B encodes the non-receptor tyrosine kinase PYK2 (protein tyrosine kinase 2 beta), a Ca2+- and integrin-responsive signaling node that links extracellular cues to cytoskeletal remodeling and gene expression. PYK2 participates in focal adhesion and actin dynamics pathways and interfaces with Src-family kinases, MAPK/ERK signaling, and downstream transcriptional programs that regulate adhesion, migration, and inflammatory responses. In immune and myeloid lineages, PYK2 modulates chemokine and cytokine-driven signaling and contributes to innate immune activation and leukocyte trafficking. Dysregulated PTK2B/PYK2 signaling has been implicated in neuroinflammatory and neurodegenerative mechanisms, autoimmune-associated signaling networks, and tumor cell invasion contexts, making it a useful target for pathway perturbation studies.

PYK2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTK2B expression without altering the underlying DNA sequence.

PYK2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTK2B 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 PTK2B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PYK2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTK2B locus and enabling the study of PYK2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PYK2 pathway restoration in tumor cells with silenced or reduced PTK2B expression.

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