CGRP2 CRISPR Activation Plasmid (h): sc-401165-ACT

Human CALCB encodes calcitonin gene-related peptide 2 (CGRP2), a neuropeptide produced from the calcitonin gene family that signals through the CGRP receptor complex (CALCRL with RAMP1) to regulate cAMP-dependent pathways. CGRP2 contributes to neurovascular communication by modulating vasodilation, vascular tone, and inflammatory neurogenic signaling, and it can influence smooth muscle and immune cell responses in peripheral tissues. Dysregulated CGRP-family signaling has been associated with altered nociceptive processing and vascular homeostasis, making CALCB a relevant target for studying sensory neuron biology, endothelial–neuronal cross-talk, and inflammatory pain mechanisms. In cancer and other chronic conditions, changes in neuropeptide signaling may shape the tissue microenvironment and stress-response pathways, supporting investigation of CALCB in cell–cell communication models.

CGRP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CALCB expression without altering the underlying DNA sequence.

CGRP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CALCB 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 CALCB transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CGRP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CALCB locus and enabling the study of CGRP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CGRP2 pathway restoration in tumor cells with silenced or reduced CALCB expression.

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