TRPC5 CRISPR Activation Plasmid (h): sc-403745-ACT

TRPC5 encodes a non-selective, Ca2+-permeable cation channel of the canonical transient receptor potential (TRPC) family that contributes to receptor-operated calcium entry at the plasma membrane. TRPC5 integrates signals downstream of G protein-coupled receptors and receptor tyrosine kinases through phospholipase C-dependent pathways, shaping intracellular Ca2+ dynamics that regulate membrane excitability, cytoskeletal remodeling, and transcriptional responses. Channel activity has been linked to neuronal and sensory signaling, vascular and renal physiology, and modulation of cell migration. Dysregulated TRPC5 signaling is frequently studied in the context of pain and anxiety-related behaviors, podocyte injury and proteinuric kidney disease mechanisms, and cancer cell motility and drug response phenotypes.

TRPC5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRPC5 expression without altering the underlying DNA sequence.

TRPC5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRPC5 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 TRPC5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRPC5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRPC5 locus and enabling the study of TRPC5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRPC5 pathway restoration in tumor cells with silenced or reduced TRPC5 expression.

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