C1orf68 CRISPR Activation Plasmid (h): sc-403506-ACT

Human C1orf68 (chromosome 1 open reading frame 68) encodes a poorly characterized protein with limited functional annotation and emerging evidence of context-dependent expression across tissues and disease states. Although specific molecular pathways remain to be fully resolved, transcript-level studies suggest C1orf68 may participate in cellular programs linked to differentiation, stress responses, and regulation of gene expression networks. Altered expression patterns have been reported in multiple omics datasets, supporting use as a candidate gene for mechanistic studies of signaling and transcriptional control in disease-relevant cellular models. Investigating C1orf68 can help clarify how understudied open reading frame genes contribute to pathway remodeling and phenotypic variation.

C1orf68 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous C1orf68 expression without altering the underlying DNA sequence.

C1orf68 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the C1orf68 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 C1orf68 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous C1orf68 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native C1orf68 locus and enabling the study of C1orf68-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of C1orf68 pathway restoration in tumor cells with silenced or reduced C1orf68 expression.

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