Dia 2 CRISPR Activation Plasmid (h): sc-403238-ACT

DIAPH2 (Dia 2) encodes a diaphanous-related formin that regulates actin filament nucleation and elongation, contributing to cytoskeletal remodeling, cell polarity, and cytokinesis. As an effector in Rho GTPase–linked signaling, DIAPH2 supports processes such as cell migration and adhesion through coordination of actin dynamics with membrane trafficking and microtubule crosstalk. Altered DIAPH2 activity and dosage have been investigated in contexts of reproductive biology and tumor cell behavior, where cytoskeletal control influences chromosomal stability and invasive phenotypes. In biomedical research, DIAPH2 is commonly studied to connect Rho–formin signaling to morphogenesis, cell-cycle progression, and mechanotransduction.

Dia 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DIAPH2 expression without altering the underlying DNA sequence.

Dia 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DIAPH2 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 DIAPH2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Dia 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DIAPH2 locus and enabling the study of Dia 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Dia 2 pathway restoration in tumor cells with silenced or reduced DIAPH2 expression.

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