HCN2 CRISPR Activation Plasmid (h): sc-405074-ACT

HCN2 encodes the hyperpolarization-activated cyclic nucleotide–gated channel 2, a pore-forming subunit that conducts the Ih current and links membrane hyperpolarization to depolarizing inward cation flux. By integrating voltage sensing with direct modulation by cyclic nucleotides, HCN2 helps tune neuronal excitability, pacemaker activity, and rhythmic firing properties, influencing signal integration and network oscillations. HCN2 function intersects with cAMP/PKA-linked signaling and broader electrophysiological programs that shape synaptic responsiveness and excitability-dependent gene regulation. Dysregulated HCN2 expression or channel activity has been associated with altered excitability phenotypes relevant to neurological and sensory system disorders, supporting mechanistic studies in excitable tissues and disease-relevant cellular models.

HCN2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HCN2 expression without altering the underlying DNA sequence.

HCN2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HCN2 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 HCN2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HCN2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HCN2 locus and enabling the study of HCN2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HCN2 pathway restoration in tumor cells with silenced or reduced HCN2 expression.

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