TRPC6 CRISPR Activation Plasmid (m): sc-423517-ACT

Trpc6 encodes TRPC6, a nonselective, calcium-permeable cation channel that functions downstream of phospholipase C signaling to couple receptor stimulation with intracellular Ca²⁺ influx. In mouse cells, TRPC6 participates in regulation of membrane excitability, cytoskeletal remodeling, and mechanosensitive responses, integrating cues from G protein–coupled and tyrosine kinase receptors. Channel activity influences Ca²⁺-dependent pathways including calcineurin/NFAT and MAPK signaling, shaping transcriptional programs and cellular contractility. Dysregulated TRPC6 signaling has been linked to kidney glomerular biology and podocyte function as well as vascular and cardiac remodeling processes, making it relevant for studying calcium-driven pathophysiology.

TRPC6 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Trpc6 expression without altering the underlying DNA sequence.

TRPC6 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Trpc6 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 Trpc6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRPC6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Trpc6 locus and enabling the study of TRPC6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRPC6 pathway restoration in tumor cells with silenced or reduced Trpc6 expression.

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