alpha Synuclein CRISPR Activation Plasmid (h): sc-417273-ACT

SNCA encodes human alpha Synuclein, a presynaptic protein implicated in synaptic vesicle trafficking, neurotransmitter release, and regulation of membrane curvature through lipid binding. It associates with vesicle recycling machinery and influences proteostasis pathways, including ubiquitin–proteasome and autophagy-lysosome systems that control protein turnover and aggregation. Dysregulated SNCA expression or misfolding is linked to neuronal vulnerability and has been extensively studied in the context of Lewy body pathology and Parkinson’s disease-related mechanisms. As a modulator of synaptic homeostasis and intracellular trafficking, SNCA is frequently used to interrogate stress responses, mitochondrial dysfunction, and neuroinflammatory signaling in cellular models.

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

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

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