E6BP CRISPR Activation Plasmid (h): sc-409673-ACT

RCN2 encodes the human E6-binding protein (E6BP), an endoplasmic reticulum–resident, calcium-binding protein implicated in secretory pathway homeostasis. E6BP contributes to calcium-dependent protein folding and quality control within the ER lumen, linking it to ER stress signaling and proteostasis-associated processes. Through these functions, RCN2 is relevant to cellular responses that modulate trafficking, secretion, and stress-adaptive transcriptional programs. Altered ER proteostasis and calcium handling are frequently associated with disease-relevant phenotypes, making RCN2 a useful target for mechanistic studies of stress signaling and secretory biology.

E6BP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RCN2 expression without altering the underlying DNA sequence.

E6BP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RCN2 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 RCN2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous E6BP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RCN2 locus and enabling the study of E6BP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of E6BP pathway restoration in tumor cells with silenced or reduced RCN2 expression.

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