Neuregulin-2 CRISPR Activation Plasmid (h): sc-404043-ACT

NRG2 encodes neuregulin-2, a member of the EGF-like neuregulin family that functions as a ligand for ERBB receptor tyrosine kinases. Through ERBB2/ERBB3 and ERBB4 signaling, Neuregulin-2 modulates neuronal and glial communication, synapse development, and activity-dependent plasticity, while also influencing PI3K–AKT and MAPK pathway outputs that regulate survival and differentiation. NRG2 expression and ERBB pathway dynamics have been linked to neurodevelopmental and neuropsychiatric phenotypes, and altered neuregulin–ERBB signaling is studied in the context of circuit dysfunction and synaptic imbalance. These properties make NRG2 a useful target for dissecting receptor-ligand signaling, transcriptional networks, and cell-type–specific responses in human cellular models.

Neuregulin-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NRG2 expression without altering the underlying DNA sequence.

Neuregulin-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NRG2 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 NRG2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Neuregulin-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NRG2 locus and enabling the study of Neuregulin-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Neuregulin-2 pathway restoration in tumor cells with silenced or reduced NRG2 expression.

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