nm23-H7 CRISPR Activation Plasmid (h): sc-403079-ACT

Human NME7 encodes nm23-H7, a member of the nucleoside diphosphate kinase (NDPK/NME) family implicated in regulation of nucleotide homeostasis and signaling outputs that depend on balanced NTP pools. nm23-H7 has been linked to centrosome- and microtubule-associated processes, supporting cell-cycle progression, intracellular transport, and organization of cytoskeletal architecture. Altered NME family activity has been observed across multiple disease contexts, making NME7 a useful node for studying how perturbations in nucleotide metabolism intersect with proliferative signaling and genome maintenance. As a result, NME7 is frequently examined in models of aberrant cell growth, chromosomal instability, and stress-adaptive transcriptional programs.

nm23-H7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NME7 expression without altering the underlying DNA sequence.

nm23-H7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NME7 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 NME7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous nm23-H7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NME7 locus and enabling the study of nm23-H7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of nm23-H7 pathway restoration in tumor cells with silenced or reduced NME7 expression.

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