RGC32 CRISPR Activation Plasmid (h): sc-404843-ACT

RGCC encodes response gene to complement 32 (RGC32), a cell cycle- and inflammation-associated protein induced by complement activation and diverse cellular stress signals. RGC32 has been linked to regulation of G2/M progression, DNA replication dynamics, and cytoskeletal remodeling through interactions with cyclin-dependent kinase pathways and Rho family signaling that influence cell migration. In vascular and epithelial contexts, RGC32 is implicated in modulating TGF‑β–driven responses, oxidative stress programs, and immune signaling, processes frequently investigated in fibrosis, vascular remodeling, and tumor microenvironment biology. Altered RGCC expression has been reported across multiple disease settings, supporting its use as a mechanistic node for studies of proliferation, differentiation, and inflammatory gene networks.

RGC32 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RGCC expression without altering the underlying DNA sequence.

RGC32 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RGCC 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 RGCC transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RGC32 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RGCC locus and enabling the study of RGC32-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RGC32 pathway restoration in tumor cells with silenced or reduced RGCC expression.

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