CRLR CRISPR Activation Plasmid (h): sc-401631-ACT

CALCRL encodes calcitonin receptor-like receptor (CRLR), a class B GPCR that forms functional receptor complexes with RAMP proteins to mediate signaling by vasoactive peptides such as CGRP and adrenomedullin. Upon ligand engagement, CRLR activates G protein–coupled pathways including cAMP/PKA and downstream transcriptional programs that regulate vascular tone, endothelial barrier function, lymphatic biology, and inflammatory responses. CRLR activity intersects with angiogenic and stress-response networks, influencing processes such as vasodilation, fluid homeostasis, and neurovascular signaling. Dysregulated CALCRL expression or signaling has been associated with cardiovascular and lymphatic phenotypes and has been studied in contexts of migraine-related neurovascular pathways, pulmonary vascular remodeling, and tumor-associated angiogenesis.

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

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

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