HIPK2 CRISPR Activation Plasmid (h): sc-402003-ACT

Human HIPK2 (homeodomain-interacting protein kinase 2) is a serine/threonine kinase that integrates stress and developmental cues to regulate transcription, apoptosis, DNA damage responses, and cell-cycle checkpoints. It modulates multiple signaling networks, including p53-dependent transcriptional programs, TGF-β/SMAD signaling, and Wnt/β-catenin pathway outputs through phosphorylation of transcription factors and co-regulators. HIPK2 activity influences chromatin-associated gene regulation and can impact cellular senescence and differentiation. Dysregulated HIPK2 expression or signaling has been linked to altered tumor suppressor pathways and aberrant stress responses, making it a relevant target for mechanistic studies in cancer biology and related cellular stress models.

HIPK2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HIPK2 expression without altering the underlying DNA sequence.

HIPK2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HIPK2 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 HIPK2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HIPK2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HIPK2 locus and enabling the study of HIPK2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HIPK2 pathway restoration in tumor cells with silenced or reduced HIPK2 expression.

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