Endophilin III CRISPR Activation Plasmid (h): sc-403821-ACT

SH3GL3 encodes endophilin III, a BAR domain-containing membrane remodeling protein that participates in clathrin-mediated endocytosis and synaptic vesicle recycling by sensing and generating membrane curvature. Endophilin III coordinates with dynamin and other endocytic adaptors to promote vesicle budding and trafficking, linking membrane dynamics to cytoskeletal organization and receptor internalization. SH3GL3 expression is enriched in neuronal tissues and is commonly studied in pathways governing neurotransmission, vesicle turnover, and intracellular signaling. Altered endocytic regulation and synaptic vesicle cycling are frequently implicated in neurobiology-related disease mechanisms, making SH3GL3 a useful target for mechanistic studies of neuronal function and membrane trafficking defects.

Endophilin III CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SH3GL3 expression without altering the underlying DNA sequence.

Endophilin III CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SH3GL3 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 SH3GL3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Endophilin III expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SH3GL3 locus and enabling the study of Endophilin III-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Endophilin III pathway restoration in tumor cells with silenced or reduced SH3GL3 expression.

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