HRC CRISPR Activation Plasmid (m): sc-420947-ACT

Mouse Hrc encodes histidine-rich calcium-binding protein (HRC), a luminal component of the sarcoplasmic reticulum that modulates Ca²⁺ storage and release by influencing ryanodine receptor activity and SERCA-dependent reuptake. By shaping excitation–contraction coupling and intracellular calcium homeostasis, HRC contributes to cardiomyocyte and skeletal muscle function, where precise Ca²⁺ cycling governs contractility and stress responses. Dysregulated Hrc expression or altered Ca²⁺ handling is commonly studied in the context of cardiac remodeling, arrhythmogenic mechanisms, and myopathy-associated phenotypes. As a nodal regulator of SR calcium dynamics, HRC is relevant to pathways controlling calcium signaling, proteostasis, and muscle adaptation to physiological and pathological stimuli.

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

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

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