Tachykinin CRISPR Activation Plasmid (h): sc-400690-ACT

TAC1 encodes tachykinin precursor peptides that are processed to neuroactive ligands such as substance P and neurokinin A, which signal primarily through neurokinin receptors to regulate neurotransmission, nociception, neurogenic inflammation, and smooth muscle tone. Tachykinin signaling engages G protein–coupled receptor pathways including phospholipase C/IP3–Ca²⁺ flux, PKC activation, and MAPK/ERK cascades, influencing cytokine release and neuronal excitability. In human tissues, TAC1 expression is enriched in sensory neurons and selected neuroendocrine and immune contexts, linking it to inflammatory circuitry and neuroimmune cross-talk. Dysregulated tachykinin pathways have been associated in the literature with chronic pain states, airway hyperreactivity, gastrointestinal motility disorders, and neuroinflammatory processes, supporting mechanistic studies of pathway modulation.

Tachykinin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TAC1 expression without altering the underlying DNA sequence.

Tachykinin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TAC1 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 TAC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Tachykinin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TAC1 locus and enabling the study of Tachykinin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Tachykinin pathway restoration in tumor cells with silenced or reduced TAC1 expression.

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