Neurogenin 3 CRISPR Activation Plasmid (h): sc-403619-ACT

NEUROG3 (Neurogenin 3) is a basic helix–loop–helix transcription factor that acts as a master regulator of endocrine lineage specification, particularly during pancreatic development where it promotes endocrine progenitor commitment and differentiation. By binding E-box motifs and coordinating transcriptional networks with pathways such as Notch-mediated lateral inhibition, NEUROG3 helps control the balance between progenitor maintenance and endocrine fate decisions. Its activity influences the emergence of insulin-, glucagon-, and somatostatin-producing cell programs, making it central to studies of pancreatic islet formation and endocrine cell identity. Altered NEUROG3 function or expression is linked to congenital disorders of enteroendocrine and pancreatic endocrine development and is frequently investigated in models of diabetes-related β-cell insufficiency and gastrointestinal endocrine dysfunction.

Neurogenin 3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NEUROG3 expression without altering the underlying DNA sequence.

Neurogenin 3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NEUROG3 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 NEUROG3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Neurogenin 3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NEUROG3 locus and enabling the study of Neurogenin 3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Neurogenin 3 pathway restoration in tumor cells with silenced or reduced NEUROG3 expression.

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