Capicua CRISPR Activation Plasmid (h): sc-406191-ACT

Human CIC encodes Capicua, an HMG-box DNA-binding transcriptional repressor that integrates receptor tyrosine kinase–RAS–MAPK/ERK signaling into context-dependent gene expression programs. Capicua constrains expression of ETS and other immediate-early transcriptional targets, shaping proliferation, differentiation, and lineage commitment by coupling extracellular cues to chromatin-associated repression. Disruption or attenuation of CIC-mediated repression has been linked to altered MAPK output and transcriptional dysregulation observed in multiple tumor contexts, where CIC can function as a molecular brake on oncogenic signaling. As a result, CIC is frequently studied in signaling-dependent transcription, cell state transitions, and mechanisms of pathway-driven gene regulatory rewiring.

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

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

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