N-Myc CRISPR Activation Plasmid (h): sc-400253-ACT

MYCN encodes the human N-Myc transcription factor, a basic helix–loop–helix leucine zipper protein that heterodimerizes with MAX to bind E-box regulatory elements and coordinate broad gene expression programs. N-Myc helps regulate cell-cycle progression, ribosome biogenesis, metabolism, DNA replication stress responses, and differentiation state through integration with MYC/MAX network activity and chromatin-associated cofactor recruitment. Dysregulated MYCN expression is linked to altered proliferative capacity and lineage programs in multiple tumor contexts, making it a widely used node for studying oncogenic transcriptional circuitry. Its downstream effects on RNA polymerase II transcription, chromatin accessibility, and growth-factor signaling pathways provide measurable readouts for mechanistic research.

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

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

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