CRHSP-24 CRISPR Activation Plasmid (h): sc-406630-ACT

CARHSP1 encodes CRHSP-24, a calcium-regulated phosphoprotein that integrates Ca2+/calmodulin-dependent signaling with post-transcriptional gene control. CRHSP-24 has been linked to modulation of mRNA stability and translation through interactions with RNA-binding protein complexes, aligning it with pathways that tune cellular stress responses and stimulus-coupled protein synthesis. As a substrate for phosphorylation-dependent regulation, it provides a mechanistic entry point for studying how kinase and phosphatase networks reshape gene expression programs. Dysregulation of calcium signaling and RNA metabolism is a recurring feature across inflammation, metabolic dysfunction, and cancer-associated phenotypes, making CARHSP1 a relevant molecular node for pathway-focused research.

CRHSP-24 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CARHSP1 expression without altering the underlying DNA sequence.

CRHSP-24 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CARHSP1 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 CARHSP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CRHSP-24 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CARHSP1 locus and enabling the study of CRHSP-24-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CRHSP-24 pathway restoration in tumor cells with silenced or reduced CARHSP1 expression.

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