Nova-2 CRISPR Activation Plasmid (h): sc-404452-ACT

Human NOVA2 encodes Nova-2, a neuron-enriched RNA-binding protein that recognizes YCAY motifs to regulate alternative splicing and other post-transcriptional RNA processing events. Nova-2 shapes neuronal differentiation and synaptic function by coordinating exon inclusion and exclusion across networks of transcripts involved in neurite outgrowth, cytoskeletal dynamics, and neurotransmission. Through these splicing programs, NOVA2 contributes to the maintenance of neural circuit integrity and activity-dependent gene regulation. Dysregulation or pathogenic variation in NOVA2 has been linked to neurodevelopmental phenotypes and neurological dysfunction, making it a useful target for mechanistic studies of RNA processing in the nervous system.

Nova-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NOVA2 expression without altering the underlying DNA sequence.

Nova-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NOVA2 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 NOVA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nova-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NOVA2 locus and enabling the study of Nova-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nova-2 pathway restoration in tumor cells with silenced or reduced NOVA2 expression.

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