MATH-2 CRISPR Activation Plasmid (h): sc-404019-ACT

NEUROD6 (MATH-2) is a basic helix–loop–helix transcription factor that regulates neuronal lineage commitment, differentiation, and maturation programs during human neurodevelopment. By binding E-box motifs, MATH-2 coordinates gene expression networks controlling neurite outgrowth, synaptic function, and neuronal survival, integrating with developmental transcriptional cascades that shape cortical and hippocampal circuitry. NEUROD6 activity is linked to cell fate decisions and excitability-related pathways in neurons, making it relevant for studying mechanisms underlying neurodevelopmental and neuropsychiatric disease biology. Perturbation of NEUROD6-regulated transcription can alter neuronal identity and connectivity, supporting its use in modeling gene regulatory networks in human neural cells.

MATH-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NEUROD6 expression without altering the underlying DNA sequence.

MATH-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NEUROD6 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 NEUROD6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MATH-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NEUROD6 locus and enabling the study of MATH-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MATH-2 pathway restoration in tumor cells with silenced or reduced NEUROD6 expression.

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