SCAMP1 CRISPR Activation Plasmid (h): sc-403757-ACT

SCAMP1 (secretory carrier membrane protein 1) is a tetraspan membrane protein enriched on recycling endosomes and transport vesicles, where it supports regulated membrane trafficking, vesicle budding, and cargo recycling to the plasma membrane. It functions within endocytic and exocytic pathways that coordinate receptor turnover, synaptic vesicle dynamics, and secretory granule release, linking SCAMP1 activity to cellular homeostasis and signaling responsiveness. By influencing vesicle-mediated transport and membrane remodeling, SCAMP1 can modulate surface availability of signaling receptors and transporters, processes frequently perturbed in proliferative and neurobiological disorders. Dysregulated trafficking networks involving SCAMP family proteins have been investigated in contexts including altered secretion, aberrant receptor recycling, and changes in cell migration and invasion-related phenotypes.

SCAMP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SCAMP1 expression without altering the underlying DNA sequence.

SCAMP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SCAMP1 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 SCAMP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SCAMP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SCAMP1 locus and enabling the study of SCAMP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SCAMP1 pathway restoration in tumor cells with silenced or reduced SCAMP1 expression.

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