nm23-H1 CRISPR Activation Plasmid (h): sc-401221-ACT

NME1 encodes nm23-H1, a nucleoside diphosphate kinase that maintains cellular NTP/NDP balance and contributes to signaling processes linked to proliferation, motility, and stress responses. nm23-H1 has been implicated in regulation of cytoskeletal dynamics, membrane trafficking, and transcriptional programs that shape cell migration and differentiation. Altered NME1 expression is frequently studied in the context of tumor biology, where it has been associated with invasion and metastatic behavior in a cell type- and context-dependent manner. As a broadly expressed human gene, NME1 provides a useful node for interrogating metabolic–signaling coupling and phenotype plasticity in engineered cellular models.

nm23-H1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NME1 expression without altering the underlying DNA sequence.

nm23-H1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NME1 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 NME1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous nm23-H1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NME1 locus and enabling the study of nm23-H1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of nm23-H1 pathway restoration in tumor cells with silenced or reduced NME1 expression.

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