C9orf152 CRISPR Activation Plasmid (h2): sc-415979-ACT-2

Human C9orf152 encodes a largely uncharacterized protein with emerging evidence for roles in cellular homeostasis and regulatory processes, supported by expression patterns and co-expression networks that implicate nuclear and cytoplasmic functions. Transcriptomic analyses suggest potential involvement in signaling and gene-expression control pathways linked to cell cycle regulation and stress responses, making it a useful target for dissecting poorly annotated regulatory nodes. Altered C9orf152 expression has been reported across disease-associated datasets, including cancer-related profiles, indicating relevance for mechanistic studies of dysregulated transcriptional programs and cellular adaptation. Gene editing of C9orf152 enables functional interrogation through knockout or precise variant modeling to map downstream pathway effects, protein interaction networks, and phenotype-genotype relationships in human cell systems.

C9orf152 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous C9orf152 expression without altering the underlying DNA sequence.

C9orf152 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the C9orf152 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 C9orf152 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous C9orf152 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native C9orf152 locus and enabling the study of C9orf152-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of C9orf152 pathway restoration in tumor cells with silenced or reduced C9orf152 expression.

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