RSAD2 CRISPR Activation Plasmid (h): sc-402884-ACT

RSAD2 (viperin) is an interferon-stimulated antiviral effector that localizes to the endoplasmic reticulum and lipid droplets and is rapidly induced downstream of pattern-recognition receptor signaling. It participates in innate immune pathways coordinated by type I interferons and NF-κB, shaping cellular responses to viral infection by modulating membrane-associated processes and immunometabolism. RSAD2 has been linked to regulation of inflammatory signaling and cytokine output, and its expression serves as a molecular readout of interferon pathway engagement. Dysregulated RSAD2 activity and interferon signatures are frequently studied in the context of chronic viral infection and interferon-associated immune pathology.

RSAD2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RSAD2 expression without altering the underlying DNA sequence.

RSAD2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RSAD2 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 RSAD2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RSAD2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RSAD2 locus and enabling the study of RSAD2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RSAD2 pathway restoration in tumor cells with silenced or reduced RSAD2 expression.

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