C18orf34 CRISPR Activation Plasmid (h): sc-415693-ACT

CCDC178 encodes the human protein C18orf34, a coiled-coil–containing factor with limited functional annotation that is predicted to participate in protein–protein interaction networks and subcellular organization. Coiled-coil proteins commonly contribute to cytoskeletal dynamics, centrosomal processes, vesicle trafficking, and assembly of multi-protein complexes, making C18orf34 relevant to studies of cell architecture and signal coordination. Emerging transcriptomic and proteomic datasets implicate C18orf34 expression changes in disease-associated cellular states, including altered proliferation and stress-response programs, although direct mechanistic links remain to be defined. These features make CCDC178 a useful target for interrogating context-dependent gene regulation and downstream pathway remodeling in human cell models.

C18orf34 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCDC178 expression without altering the underlying DNA sequence.

C18orf34 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCDC178 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 CCDC178 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous C18orf34 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCDC178 locus and enabling the study of C18orf34-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of C18orf34 pathway restoration in tumor cells with silenced or reduced CCDC178 expression.

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