β2-Adaptin CRISPR Activation Plasmid (h): sc-402404-ACT

AP2B1 encodes the human β2-Adaptin subunit of the adaptor protein 2 (AP-2) complex, a core component of clathrin-mediated endocytosis at the plasma membrane. By coupling cargo recognition motifs to clathrin coat assembly, β2-Adaptin supports internalization and recycling of receptors, transporters, and signaling complexes, thereby shaping membrane trafficking, synaptic vesicle cycling, and signal attenuation. AP-2–dependent trafficking influences pathways such as receptor tyrosine kinase and GPCR signaling, immune receptor turnover, and nutrient uptake through endocytic sorting decisions. Dysregulation of endocytic adaptor function has been linked to neurodevelopmental and neuropsychiatric phenotypes, altered synaptic function, and broader cellular vulnerability to proteostasis and trafficking stress.

β2-Adaptin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AP2B1 expression without altering the underlying DNA sequence.

β2-Adaptin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AP2B1 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 AP2B1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous β2-Adaptin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AP2B1 locus and enabling the study of β2-Adaptin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of β2-Adaptin pathway restoration in tumor cells with silenced or reduced AP2B1 expression.

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