IP3R-II CRISPR Activation Plasmid (m): sc-421193-ACT

Mouse Itpr2 encodes inositol 1,4,5-trisphosphate receptor type 2 (IP3R-II), an endoplasmic reticulum Ca²⁺-release channel that converts IP3 signals into cytosolic calcium transients. IP3R-II functions downstream of GPCR and receptor tyrosine kinase pathways that stimulate phospholipase C, shaping Ca²⁺-dependent processes such as secretion, contraction, metabolism, and transcriptional responses. By regulating spatiotemporal Ca²⁺ dynamics and ER Ca²⁺ homeostasis, Itpr2 influences inter-organelle signaling and stress responses that are frequently perturbed in pathophysiology. Dysregulated IP3R-mediated calcium signaling has been implicated in diverse disease-relevant mechanisms, including altered excitability, epithelial transport defects, and disrupted cellular homeostasis in neurological and endocrine contexts.

IP3R-II CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Itpr2 expression without altering the underlying DNA sequence.

IP3R-II CRISPR Activation Plasmid (m) 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.