C14orf37 CRISPR Activation Plasmid (h): sc-414550-ACT

C14orf37 (human) encodes a poorly characterized protein with limited functional annotation, and current evidence suggests it may contribute to context-dependent regulation of gene expression and cellular homeostasis. Transcript-level studies indicate that C14orf37 expression can vary across tissues and experimental perturbations, making it a candidate locus for investigating transcriptional control and cell state transitions. Although definitive pathway placement remains incomplete, emerging associations from genomics and expression profiling link C14orf37 to biological programs relevant to proliferation, differentiation, and stress-responsive signaling. As a result, C14orf37 is of interest for mechanistic studies that connect endogenous gene dosage with phenotypes observed in disease-relevant cellular models.

C14orf37 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous C14orf37 expression without altering the underlying DNA sequence.

C14orf37 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the C14orf37 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 C14orf37 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous C14orf37 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native C14orf37 locus and enabling the study of C14orf37-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of C14orf37 pathway restoration in tumor cells with silenced or reduced C14orf37 expression.

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