parvalbumin α CRISPR Activation Plasmid (h): sc-401364-ACT

PVALB encodes parvalbumin alpha, a high-affinity EF-hand calcium-binding protein that shapes intracellular Ca²⁺ dynamics by accelerating Ca²⁺ buffering and relaxation in excitable cells. In the nervous system, parvalbumin marks fast-spiking GABAergic interneurons and supports precise spike timing and network oscillations by influencing Ca²⁺-dependent neurotransmitter release and synaptic integration. Beyond neural circuits, calcium buffering by parvalbumin contributes to contraction–relaxation kinetics in muscle and other Ca²⁺-regulated cellular processes. Altered PVALB expression and parvalbumin-positive interneuron function have been associated with neurodevelopmental and neuropsychiatric phenotypes, making PVALB regulation relevant to studies of circuit excitability and calcium-dependent signaling.

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

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

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