BPAG1 CRISPR Activation Plasmid (h): sc-403388-ACT

DST encodes bullous pemphigoid antigen 1 (BPAG1), a large cytolinker protein that connects intermediate filaments to membrane-associated complexes and supports cytoskeletal organization, vesicle transport, and cellular mechanical resilience. Human BPAG1 isoforms are prominent in stratified epithelia and neurons, where they help maintain hemidesmosome integrity and stabilize cell–matrix adhesion. Through these structural roles, BPAG1 influences processes such as migration, polarity, and stress-response signaling that depend on coordinated actin–microtubule–intermediate filament crosstalk. Dysregulation or altered antigenicity of BPAG1 is associated with blistering skin pathology and neurodegenerative phenotypes, making DST a useful target for studying cytoskeletal dysfunction and tissue fragility mechanisms.

BPAG1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DST expression without altering the underlying DNA sequence.

BPAG1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DST 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 DST transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BPAG1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DST locus and enabling the study of BPAG1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BPAG1 pathway restoration in tumor cells with silenced or reduced DST expression.

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