NT-4 CRISPR Activation Plasmid (h): sc-402291-ACT

Human NTF4 encodes neurotrophin-4 (NT-4), a secreted growth factor that signals primarily through NTRK2/TrkB and, with lower affinity, NGFR/p75NTR to regulate neuronal survival, differentiation, and synaptic maintenance. NT-4–TrkB engagement activates downstream PI3K–AKT, MAPK/ERK, and PLCγ signaling, supporting neurite outgrowth and activity-dependent plasticity across central and peripheral nervous systems. Beyond neurobiology, NTF4 expression and TrkB pathway activity influence cellular stress responses and trophic support programs that intersect with degeneration and maladaptive remodeling. Dysregulated neurotrophin/TrkB signaling has been studied in contexts including neurodegenerative processes, neuropathic pain mechanisms, and tumor cell–microenvironment interactions where growth factor availability can shape proliferation and survival phenotypes.

NT-4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NTF4 expression without altering the underlying DNA sequence.

NT-4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NTF4 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 NTF4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NT-4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NTF4 locus and enabling the study of NT-4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NT-4 pathway restoration in tumor cells with silenced or reduced NTF4 expression.

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