γC-crystallin CRISPR Activation Plasmid (h): sc-402940-ACT

CRYGC encodes human γC-crystallin, a highly abundant structural protein of the ocular lens that contributes to refractive index and long-term transparency through stable, tightly packed protein assemblies. As a member of the β/γ-crystallin superfamily, γC-crystallin supports lens fiber cell architecture and proteostasis under conditions of minimal protein turnover. Disruption of crystallin homeostasis can promote protein misfolding and aggregation, linking CRYGC dysregulation to lens opacity phenotypes. CRYGC is therefore widely used as a marker and mechanistic entry point for studying lens development, stress responses, and protein aggregation processes relevant to cataract biology.

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

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

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