VILIP-1 CRISPR Activation Plasmid (h): sc-401100-ACT

VSNL1 encodes visinin-like protein 1 (VILIP-1), a neuronal calcium sensor of the EF-hand family that undergoes Ca2+-dependent conformational changes to regulate signaling at membranes. VILIP-1 participates in Ca2+-coupled modulation of synaptic activity and neuronal excitability, influencing second-messenger pathways and downstream transcriptional programs important for neuroplasticity. Altered VSNL1/VILIP-1 expression and calcium signaling dynamics have been associated with neurodegenerative and neuropsychiatric disease-relevant processes, including neuronal stress responses and synaptic dysfunction. Accordingly, VSNL1 is frequently studied in models of neuronal differentiation, activity-dependent signaling, and biomarker-linked molecular phenotypes.

VILIP-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous VSNL1 expression without altering the underlying DNA sequence.

VILIP-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the VSNL1 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 VSNL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous VILIP-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native VSNL1 locus and enabling the study of VILIP-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of VILIP-1 pathway restoration in tumor cells with silenced or reduced VSNL1 expression.

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