IP3R-II CRISPR Activation Plasmid (h): sc-401067-ACT

ITPR2 encodes the human inositol 1,4,5-trisphosphate receptor type 2 (IP3R-II), an ER membrane Ca2+ release channel that converts IP3 signals downstream of phospholipase C into cytosolic calcium transients. IP3R-II-mediated calcium flux shapes excitation–contraction coupling, secretion, metabolism, and transcriptional programs through Ca2+-dependent effectors such as calmodulin, CaMK, calcineurin/NFAT, and mitochondrial calcium uptake. This signaling node integrates inputs from GPCR and receptor tyrosine kinase pathways and contributes to inter-organelle calcium coupling at ER–mitochondria contact sites. Dysregulated IP3R-II activity and altered calcium homeostasis have been implicated in mechanistic studies of cardiovascular and epithelial physiology and in research on disorders involving aberrant calcium signaling, including arrhythmia-associated phenotypes and proliferative or inflammatory signaling contexts.

IP3R-II CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ITPR2 expression without altering the underlying DNA sequence.

IP3R-II CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ITPR2 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 ITPR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IP3R-II expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ITPR2 locus and enabling the study of IP3R-II-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IP3R-II pathway restoration in tumor cells with silenced or reduced ITPR2 expression.

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