RICK CRISPR Activation Plasmid (h): sc-400731-ACT

Receptor-interacting serine/threonine-protein kinase 2 (RIPK2; also known as RICK) is a cytosolic signaling kinase that functions as a key adaptor downstream of NOD1 and NOD2 pattern recognition receptors. Upon sensing bacterial peptidoglycan motifs, RIPK2 mediates ubiquitin-dependent signaling to activate NF-κB and MAPK pathways, shaping inflammatory gene expression, cytokine production, and innate immune crosstalk with epithelial and myeloid cells. RIPK2 also interfaces with autophagy and cell stress signaling through regulation of ubiquitin ligases and downstream kinase cascades. Dysregulated RIPK2 pathway activity has been associated with chronic inflammatory phenotypes and immune-mediated pathology, making it a widely studied node in host–microbe interaction and inflammatory signaling research.

RICK CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RIPK2 expression without altering the underlying DNA sequence.

RICK CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RIPK2 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 RIPK2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RICK expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RIPK2 locus and enabling the study of RICK-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RICK pathway restoration in tumor cells with silenced or reduced RIPK2 expression.

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