Rtn-3 CRISPR Activation Plasmid (h): sc-404072-ACT

RTN3 encodes reticulon-3 (Rtn-3), an endoplasmic reticulum (ER) membrane protein that shapes tubular ER architecture and contributes to membrane curvature and trafficking dynamics. Rtn-3 participates in ER homeostasis, neurite outgrowth, and regulation of intracellular transport processes that influence proteostasis. In neuronal systems, RTN3 has been linked to pathways controlling amyloid precursor protein (APP) processing and axonal integrity, connecting ER stress responses and synaptic maintenance to neurodegenerative disease mechanisms. Altered RTN3 expression or localization is therefore studied in the context of ER remodeling, protein quality control, and CNS pathology-relevant signaling networks.

Rtn-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RTN3 expression without altering the underlying DNA sequence.

Rtn-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RTN3 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 RTN3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rtn-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RTN3 locus and enabling the study of Rtn-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rtn-3 pathway restoration in tumor cells with silenced or reduced RTN3 expression.

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