PIEZO1 CRISPR Activation Plasmid (h): sc-410555-ACT

PIEZO1 encodes a large mechanosensitive cation channel that converts membrane tension and shear stress into calcium influx, enabling cells to transduce mechanical cues into biochemical signals. It regulates fundamental processes including vascular tone and remodeling, erythrocyte volume control, epithelial and endothelial mechanotransduction, and immune cell activation, with downstream effects on calcium-dependent pathways such as calmodulin signaling, cytoskeletal remodeling, and YAP/TAZ-associated transcriptional programs. Altered PIEZO1 activity is linked to disorders of red blood cell hydration and morphology, and to dysregulated mechanotransduction in cardiovascular and lymphatic biology. These properties make PIEZO1 a key target for studying how mechanical forces shape cell fate decisions, barrier function, and tissue homeostasis.

PIEZO1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PIEZO1 expression without altering the underlying DNA sequence.

PIEZO1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PIEZO1 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 PIEZO1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PIEZO1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PIEZO1 locus and enabling the study of PIEZO1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PIEZO1 pathway restoration in tumor cells with silenced or reduced PIEZO1 expression.

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