LTRPC7 CRISPR Activation Plasmid (h): sc-402820-ACT

TRPM7 (LTRPC7) encodes a bifunctional channel-kinase that conducts divalent cations, particularly Mg²⁺ and Ca²⁺, and couples ion homeostasis to intracellular signaling via its serine/threonine kinase domain. TRPM7 activity influences membrane potential, cytoskeletal dynamics, and cell adhesion and migration, integrating with pathways governing proliferation, differentiation, and stress responses. Through regulation of Mg²⁺-dependent enzymatic processes and Ca²⁺-linked signaling, TRPM7 contributes to control of cellular metabolism and survival. Dysregulated TRPM7 expression or function has been associated with altered excitability and remodeling programs implicated across neurological injury models, cardiovascular pathophysiology, and cancer cell phenotypes, supporting its utility as a mechanistic target in functional genomics.

LTRPC7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRPM7 expression without altering the underlying DNA sequence.

LTRPC7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRPM7 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 TRPM7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LTRPC7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRPM7 locus and enabling the study of LTRPC7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LTRPC7 pathway restoration in tumor cells with silenced or reduced TRPM7 expression.

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