beta Synuclein CRISPR Activation Plasmid (h): sc-400894-ACT

Human SNCB encodes beta Synuclein, a presynaptic, cytosolic protein that modulates synaptic vesicle dynamics, neurotransmitter release, and membrane-associated protein trafficking in neurons. As a member of the synuclein family, it participates in protein–lipid interactions and influences proteostasis pathways that govern aggregation, turnover, and stress responses at the synapse. Altered synuclein biology is closely linked to neurodegenerative processes, and SNCB is studied for its interactions with alpha-synuclein, effects on synaptic resilience, and contributions to neuronal vulnerability. Research commonly leverages SNCB to interrogate mechanisms of synaptic function, neuronal differentiation, and pathways implicated in Parkinson’s disease and related synucleinopathies.

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

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

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