REEP5 CRISPR Activation Plasmid (h): sc-405007-ACT

REEP5 (receptor expression-enhancing protein 5) is an endoplasmic reticulum–resident membrane protein that helps shape ER tubules and supports ER–plasma membrane organization, contributing to efficient trafficking and surface expression of select receptors and channels. By influencing membrane curvature and secretory pathway dynamics, REEP5 affects cellular processes including protein maturation, vesicle transport, and maintenance of ER homeostasis. Altered REEP5 activity has been linked to disrupted receptor signaling and membrane organization, making it relevant to studies of cell excitability, stress adaptation, and pathway rewiring in disease-associated states. Human REEP5 is therefore a useful target for dissecting ER morphogenesis and receptor trafficking mechanisms that intersect with signaling networks and cellular fitness.

REEP5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous REEP5 expression without altering the underlying DNA sequence.

REEP5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the REEP5 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 REEP5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous REEP5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native REEP5 locus and enabling the study of REEP5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of REEP5 pathway restoration in tumor cells with silenced or reduced REEP5 expression.

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