Amphiphysin II CRISPR Activation Plasmid (h): sc-401263-ACT

BIN1 encodes amphiphysin II, a BAR domain-containing membrane remodeling protein that coordinates clathrin-mediated endocytosis, T-tubule biogenesis, and curvature-dependent trafficking events through interactions with dynamin and phosphoinositide-rich membranes. Amphiphysin II supports vesicle formation and cytoskeletal organization, with context-dependent roles in neuronal synaptic function and muscle membrane architecture. Altered BIN1 expression and splicing have been linked to pathways relevant to myotubular myopathy and neuromuscular degeneration, and BIN1 is also studied in endocytic network changes observed in neurodegenerative disease biology. These functions make BIN1 a useful target for dissecting membrane dynamics, trafficking-dependent signaling, and cell-type-specific isoform regulation in human model systems.

Amphiphysin II CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BIN1 expression without altering the underlying DNA sequence.

Amphiphysin II CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BIN1 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 BIN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Amphiphysin II expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BIN1 locus and enabling the study of Amphiphysin II-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Amphiphysin II pathway restoration in tumor cells with silenced or reduced BIN1 expression.

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