RORγ CRISPR Activation Plasmid (h): sc-400912-ACT

RORC encodes the nuclear receptor RORγ, a ligand-regulated transcription factor that coordinates gene programs involved in immune differentiation and tissue inflammation. In human T cells, RORγt isoform activity is central to Th17 lineage commitment and regulation of cytokine networks such as IL-17A/F and IL-23–responsive pathways, linking it to host defense and mucosal barrier biology. RORγ also integrates metabolic and circadian-associated transcriptional inputs through nuclear receptor signaling and chromatin remodeling. Dysregulated RORC expression or signaling has been associated with inflammatory and autoimmune disease mechanisms and altered tumor–immune interactions, making it a useful node for pathway interrogation in immunology and cancer biology models.

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

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

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