SEC22B CRISPR Activation Plasmid (h): sc-402384-ACT

SEC22B (Sec22 homolog B) encodes a v-SNARE protein that localizes to the ER–Golgi interface and supports vesicle docking and membrane fusion within the early secretory pathway. By pairing with t-SNARE complexes, SEC22B contributes to anterograde and retrograde trafficking, maintaining protein maturation, organelle homeostasis, and secretory cargo flux. SEC22B has also been implicated in antigen cross-presentation and phagosome-to-cytosol trafficking in immune cells, connecting vesicle transport to adaptive immune signaling. Dysregulation of membrane trafficking programs involving SEC22B can influence ER stress responses and proteostasis networks that are frequently perturbed in cancer, neurodegeneration, and inflammatory phenotypes.

SEC22B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SEC22B expression without altering the underlying DNA sequence.

SEC22B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SEC22B 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 SEC22B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SEC22B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SEC22B locus and enabling the study of SEC22B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SEC22B pathway restoration in tumor cells with silenced or reduced SEC22B expression.

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