SPRR2G CRISPR Activation Plasmid (h): sc-416907-ACT

SPRR2G (small proline-rich protein 2G) is a cornified envelope precursor predominantly expressed in stratified epithelia, where it contributes to barrier formation by serving as a substrate for transglutaminase-mediated crosslinking during terminal keratinocyte differentiation. It functions within epidermal differentiation and keratinization programs coordinated with cytoskeletal remodeling and stress-responsive epithelial gene networks. Altered expression of SPRR family members is commonly observed in barrier-disruptive inflammatory skin conditions and epithelial hyperproliferation, and SPRR2G has been used as a molecular readout of differentiation state and tissue remodeling. These features make SPRR2G relevant for studying epithelial homeostasis, injury responses, and disease-associated changes in differentiation pathways.

SPRR2G CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SPRR2G expression without altering the underlying DNA sequence.

SPRR2G CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SPRR2G 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 SPRR2G transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SPRR2G expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SPRR2G locus and enabling the study of SPRR2G-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SPRR2G pathway restoration in tumor cells with silenced or reduced SPRR2G expression.

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