PADI2 CRISPR Activation Plasmid (h): sc-403349-ACT

Peptidyl arginine deiminase 2 (PADI2) is a calcium-dependent enzyme that catalyzes citrullination of protein arginine residues, altering protein charge, structure, and interaction networks. This post-translational modification regulates chromatin accessibility and gene expression through histone citrullination and can remodel cytoskeletal and myelin-associated proteins, linking PADI2 to differentiation and inflammatory signaling. PADI2 activity intersects with transcriptional programs influenced by cytokine-driven pathways and has been studied in contexts of aberrant epigenetic regulation, immune-mediated tissue injury, and tumor-associated phenotypes. Dysregulated citrullination and neoepitope generation implicate PADI2 in mechanisms relevant to autoimmunity and cancer biology, supporting its use as a molecular node for pathway interrogation.

PADI2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PADI2 expression without altering the underlying DNA sequence.

PADI2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PADI2 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 PADI2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PADI2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PADI2 locus and enabling the study of PADI2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PADI2 pathway restoration in tumor cells with silenced or reduced PADI2 expression.

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