CysLT₂ Receptor CRISPR Activation Plasmid (h): sc-403435-ACT

Human CYSLTR2 encodes the cysteinyl leukotriene receptor 2 (CysLT2 receptor), a G protein–coupled receptor that binds LTC4, LTD4, and LTE4 to regulate inflammatory signaling. Receptor activation can couple to Gq/PLCβ to elevate intracellular Ca2+, engage MAPK pathways, and modulate endothelial permeability, smooth muscle tone, and leukocyte recruitment. CYSLTR2 activity integrates into arachidonic acid and eicosanoid biology, shaping cytokine production and tissue remodeling in immune and vascular contexts. Dysregulated leukotriene receptor signaling and recurrent CYSLTR2 variants have been linked to inflammatory disease mechanisms and oncogenic signaling programs in select malignancies, supporting its utility as a pathway node in mechanistic studies.

CysLT2 Receptor CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYSLTR2 expression without altering the underlying DNA sequence.

CysLT2 Receptor CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYSLTR2 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 CYSLTR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CysLT2 Receptor expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYSLTR2 locus and enabling the study of CysLT2 Receptor-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CysLT2 Receptor pathway restoration in tumor cells with silenced or reduced CYSLTR2 expression.

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