Neurogenin 1 CRISPR Activation Plasmid (m): sc-421860-ACT

Neurog1 encodes Neurogenin 1, a basic helix–loop–helix transcription factor that functions as a master regulator of neurogenesis in the mouse nervous system. It promotes neuronal lineage commitment and differentiation by activating pro-neural transcriptional programs and coordinating cell-cycle exit, interfacing with developmental signaling networks such as Notch/Delta-mediated lateral inhibition and neurogenic bHLH factor cascades. Neurogenin 1 activity shapes sensory neuron development and cortical and spinal cord neuronal patterning, and dysregulation of neurogenic differentiation programs is relevant to studies of congenital neurodevelopmental disorders, neural tube patterning defects, and aberrant neuronal fate specification. Because it sits upstream of many neuronal subtype markers and differentiation effectors, Neurog1 is widely used to interrogate transcriptional control of neural stem/progenitor dynamics and neuronal maturation.

Neurogenin 1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Neurog1 expression without altering the underlying DNA sequence.

Neurogenin 1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Neurog1 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 Neurog1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Neurogenin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Neurog1 locus and enabling the study of Neurogenin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Neurogenin 1 pathway restoration in tumor cells with silenced or reduced Neurog1 expression.

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