Dia 1 CRISPR Activation Plasmid (h): sc-401929-ACT

DIAPH1 (Dia 1) encodes a Diaphanous-related formin that regulates actin filament nucleation and elongation downstream of Rho GTPase signaling. Dia 1 coordinates cytoskeletal remodeling required for cell polarity, adhesion, cytokinesis, and directed migration, and it also contributes to microtubule stabilization through actin–microtubule crosstalk. Through these processes, DIAPH1 influences pathways governing mechanotransduction and membrane dynamics, including filopodia formation and contractile ring assembly. Dysregulated DIAPH1 activity has been associated with altered cell motility and division phenotypes and is linked in the literature to developmental and hematologic/vascular phenotypes, supporting its relevance for disease-mechanism studies.

Dia 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DIAPH1 expression without altering the underlying DNA sequence.

Dia 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DIAPH1 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 DIAPH1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Dia 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DIAPH1 locus and enabling the study of Dia 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Dia 1 pathway restoration in tumor cells with silenced or reduced DIAPH1 expression.

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