Neuroplastin CRISPR Activation Plasmid (h): sc-404244-ACT

Human NPTN encodes neuroplastin, an immunoglobulin superfamily cell-adhesion glycoprotein enriched at neuronal synapses and also expressed in diverse non-neural tissues. Neuroplastin participates in synapse formation and stabilization, regulates excitatory neurotransmission and synaptic plasticity, and interfaces with Ca²⁺ homeostasis through functional coupling to plasma membrane Ca²⁺ ATPases, influencing downstream calcium-dependent signaling. These activities connect NPTN to pathways governing neurite outgrowth, activity-dependent circuit remodeling, and membrane protein organization. Altered neuroplastin expression or signaling has been associated with neurodevelopmental and neuropsychiatric phenotypes, and it is frequently studied in the context of cognition-related synaptic dysfunction and cell-state remodeling in disease models.

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

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

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