SNARK CRISPR Activation Plasmid (h): sc-405079-ACT

Human NUAK2 encodes SNARK (NUAK family SNF1-like kinase 2), an AMPK-related serine/threonine kinase that integrates cellular energy status with cytoskeletal remodeling, adhesion, and stress-adaptive signaling. SNARK activity has been linked to regulation of cell migration and invasion, epithelial–mesenchymal transition-associated programs, and context-dependent control of survival under metabolic stress, intersecting with pathways downstream of LKB1 and modulators of AKT/mTOR and TGF-β signaling. Altered NUAK2/SNARK expression or signaling has been reported in multiple tumor contexts and in fibrotic and inflammatory states, consistent with roles in tissue remodeling and stress responses. These properties make NUAK2 a relevant node for studying kinase-driven regulation of motility, metabolism, and transcriptional adaptation in human cell models.

SNARK CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NUAK2 expression without altering the underlying DNA sequence.

SNARK CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NUAK2 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 NUAK2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SNARK expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NUAK2 locus and enabling the study of SNARK-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SNARK pathway restoration in tumor cells with silenced or reduced NUAK2 expression.

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