WNK2 CRISPR Activation Plasmid (h): sc-407834-ACT

WNK2 (with-no-lysine [K] kinase 2) encodes a serine/threonine kinase that functions as an upstream regulator of ion transport and cell signaling, influencing phosphorylation cascades that modulate membrane transporters and cytoskeletal dynamics. In human cells, WNK2 integrates cues that affect osmotic balance and MAPK-associated signaling outputs, shaping processes such as proliferation, migration, and stress responses. Altered WNK2 expression or regulatory control has been associated with dysregulated signaling networks in cancer biology and other contexts where kinase-driven pathway balance is perturbed, making it a useful node for mechanistic studies of pathway crosstalk. As a kinase with pathway-level effects, WNK2 is frequently investigated for its impact on downstream transporter regulation and signaling rewiring in disease-relevant cellular models.

WNK2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous WNK2 expression without altering the underlying DNA sequence.

WNK2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the WNK2 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 WNK2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous WNK2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native WNK2 locus and enabling the study of WNK2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of WNK2 pathway restoration in tumor cells with silenced or reduced WNK2 expression.

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