N-type Ca CP α1B CRISPR Activation Plasmid (h): sc-402397-ACT

CACNA1B encodes the pore-forming α1B subunit of N-type voltage-gated calcium channels (CaV2.2), which mediate depolarization-evoked Ca2+ influx in excitable cells. Channel opening couples membrane activity to intracellular calcium signaling, coordinating presynaptic vesicle docking and neurotransmitter release through Ca2+-dependent pathways and downstream kinases. CACNA1B activity shapes synaptic transmission, neuronal excitability, and neuroendocrine secretion, with functional variation linked to altered pain processing and neuropsychiatric phenotypes. As a core component of calcium-dependent signaling networks, CaV2.2 is routinely studied in models of synaptic physiology and ion channel regulation.

N-type Ca CP α1B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CACNA1B expression without altering the underlying DNA sequence.

N-type Ca CP α1B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CACNA1B 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 CACNA1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous N-type Ca CP α1B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CACNA1B locus and enabling the study of N-type Ca CP α1B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of N-type Ca CP α1B pathway restoration in tumor cells with silenced or reduced CACNA1B expression.

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