p115 CRISPR Activation Plasmid (h): sc-403918-ACT

USO1 encodes the vesicle tethering factor p115, a conserved coiled-coil protein that coordinates ER-to-Golgi and intra-Golgi transport by bridging vesicle coats with Golgi matrix components and SNARE machinery. p115 supports maintenance of Golgi structure, directional membrane traffic, and secretory pathway homeostasis, thereby influencing protein processing and surface delivery. Perturbation of USO1-linked trafficking can impact proteostasis, stress responses, and signaling outputs that depend on regulated secretion and membrane receptor turnover. Dysregulated vesicular transport and Golgi organization are frequently associated with oncogenic transformation and neurodegenerative phenotypes, making USO1 a useful node for studying pathway-level consequences of secretory dysfunction.

p115 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous USO1 expression without altering the underlying DNA sequence.

p115 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the USO1 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 USO1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p115 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native USO1 locus and enabling the study of p115-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p115 pathway restoration in tumor cells with silenced or reduced USO1 expression.

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