Synaptojanin 1 CRISPR Activation Plasmid (m): sc-430506-ACT

Synj1 encodes synaptojanin 1, a polyphosphoinositide phosphatase that hydrolyzes PI(4,5)P2 and related phosphoinositides to regulate membrane trafficking. In neurons, synaptojanin 1 coordinates clathrin-mediated endocytosis and synaptic vesicle recycling by coupling phosphoinositide turnover to coat disassembly and actin remodeling. This activity intersects with endosomal sorting and autophagy-relevant membrane dynamics that shape presynaptic function and proteostasis. Dysregulated SYNJ1 activity has been linked to neurodegenerative and movement disorder biology, making it a useful node for studying synaptic maintenance pathways under stress.

Synaptojanin 1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Synj1 expression without altering the underlying DNA sequence.

Synaptojanin 1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Synj1 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 Synj1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Synaptojanin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Synj1 locus and enabling the study of Synaptojanin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Synaptojanin 1 pathway restoration in tumor cells with silenced or reduced Synj1 expression.

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