SLK CRISPR Activation Plasmid (h): sc-406490-ACT

Human SLK (STE20-like kinase) encodes a serine/threonine kinase implicated in cytoskeletal remodeling, focal adhesion turnover, and regulation of cell motility and polarity. SLK participates in signaling networks linked to MAPK-like kinase cascades and stress-responsive pathways that coordinate actin dynamics with cell-cycle progression and apoptosis. Through effects on adhesion-dependent signaling and tissue architecture, altered SLK activity has been associated with phenotypes relevant to tumor cell invasion, epithelial integrity, and developmental abnormalities. SLK is therefore frequently studied in models of migration, morphogenesis, and signaling crosstalk that couple mechanical cues to transcriptional programs.

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

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

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