Adducin α CRISPR Activation Plasmid (h): sc-401693-ACT

Human ADD1 encodes adducin alpha, a membrane skeletal protein that caps and bundles actin filaments and promotes spectrin–actin network assembly at the plasma membrane. Through regulation of cytoskeletal remodeling, cell–cell junction stability, and membrane organization, ADD1 contributes to erythrocyte integrity and broader control of cell shape and motility. Adducin phosphorylation by kinases such as PKC and Rho-associated signaling modulates its affinity for actin and spectrin, linking it to dynamic control of cortical architecture. Genetic and functional perturbations of ADD1 have been associated with altered membrane stability and signaling phenotypes that are relevant to studies of vascular biology, hypertension risk loci, and cytoskeleton-linked mechanisms in complex disease.

Adducin α CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADD1 expression without altering the underlying DNA sequence.

Adducin α CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADD1 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 ADD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Adducin α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADD1 locus and enabling the study of Adducin α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Adducin α pathway restoration in tumor cells with silenced or reduced ADD1 expression.

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