NGFR p75 CRISPR Activation Plasmid (h): sc-400552-ACT

NGFR (p75NTR) encodes the low-affinity neurotrophin receptor NGFR p75, a member of the TNF receptor superfamily that binds NGF and other neurotrophins to modulate neuronal survival, differentiation, and apoptosis in a context-dependent manner. NGFR p75 cooperates with Trk receptors and sortilin to influence ligand specificity and to regulate downstream signaling through NF-κB, JNK/c-Jun, RhoA, and ceramide-dependent pathways that shape axonal growth, synaptic remodeling, and stress responses. In human tissues, NGFR expression marks neural crest–derived lineages and dynamic injury responses, and altered p75NTR signaling has been associated with neurodegeneration, neuropathic pain mechanisms, and tumor cell plasticity in several cancers. These properties make NGFR a useful molecular handle for probing neurotrophin pathway crosstalk, cell fate transitions, and apoptosis-versus-survival signaling thresholds.

NGFR p75 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NGFR expression without altering the underlying DNA sequence.

NGFR p75 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NGFR 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 NGFR transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NGFR p75 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NGFR locus and enabling the study of NGFR p75-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NGFR p75 pathway restoration in tumor cells with silenced or reduced NGFR expression.

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