PADI4 CRISPR Activation Plasmid (h): sc-402657-ACT

PADI4 encodes peptidyl arginine deiminase 4 (PAD4), a calcium-dependent enzyme that converts arginine residues to citrulline on histones and other substrates, thereby modulating chromatin structure and transcriptional programs. Through histone citrullination, PADI4 intersects with epigenetic regulation of inflammatory gene expression, granulocyte differentiation, and innate immune responses, including pathways linked to neutrophil extracellular trap formation. Altered PADI4 activity has been associated with autoimmune and inflammatory conditions and has also been studied in the context of tumor biology where epigenetic remodeling influences cell state. These features make PADI4 a useful target for probing citrullination-dependent signaling and transcriptional control in immune and cancer-relevant models.

PADI4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PADI4 expression without altering the underlying DNA sequence.

PADI4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PADI4 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 PADI4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PADI4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PADI4 locus and enabling the study of PADI4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PADI4 pathway restoration in tumor cells with silenced or reduced PADI4 expression.

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