ELKS CRISPR Activation Plasmid (h): sc-404216-ACT

Human ERC1 encodes ELKS (also known as Rab6-interacting/CAST family scaffolding protein), a core component of the cortical active zone that organizes vesicle docking and regulated exocytosis. ELKS coordinates protein assemblies that couple small GTPase signaling with cytoskeletal and membrane trafficking pathways to support efficient neurotransmitter release and secretory granule fusion. In neurons and endocrine cells, ELKS contributes to synaptic organization, presynaptic release probability, and stimulus-dependent secretion through interactions with active zone and SNARE-associated machinery. Altered ERC1/ELKS expression or localization has been linked to dysregulated synaptic transmission and broader neurobiological phenotypes, supporting its use as a molecular handle in pathway-level studies of secretion and circuit function.

ELKS CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ERC1 expression without altering the underlying DNA sequence.

ELKS CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ERC1 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 ERC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ELKS expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ERC1 locus and enabling the study of ELKS-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ELKS pathway restoration in tumor cells with silenced or reduced ERC1 expression.

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