HUNK CRISPR Activation Plasmid (h): sc-405786-ACT

HUNK (hormonally upregulated neu-associated kinase) encodes a serine/threonine kinase implicated in signal transduction programs that influence epithelial cell behavior, including proliferation, differentiation, and survival. Reported functions connect HUNK to growth factor–responsive pathways and stress-adaptive processes that can modulate cell polarity and migratory phenotypes in a context-dependent manner. Altered HUNK expression or activity has been observed in multiple cancer-related models, supporting its relevance for studying mechanisms of oncogenic signaling and cellular plasticity. As a kinase-linked regulator, HUNK is frequently investigated for how it rewires downstream phosphorylation networks and transcriptional outputs in human cell systems.

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

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

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