TrkB CRISPR Activation Plasmid (h): sc-400142-ACT

NTRK2 encodes the human TrkB receptor tyrosine kinase, a high-affinity receptor for BDNF and related neurotrophins that controls neuronal differentiation, synaptic plasticity, and survival. Upon ligand binding, TrkB activates PI3K–AKT, MAPK/ERK, and PLCγ signaling cascades that coordinate transcriptional programs, cytoskeletal remodeling, and activity-dependent connectivity. TrkB signaling is central to neurodevelopment and circuit maintenance, and dysregulation of NTRK2 expression or pathway coupling has been linked to altered neuronal function in neuropsychiatric and neurodegenerative disease contexts. As a nodal receptor in neurotrophin pathways, NTRK2 is frequently studied for its effects on cell-state transitions, stress responses, and stimulus-driven gene expression.

TrkB CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NTRK2 expression without altering the underlying DNA sequence.

TrkB CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NTRK2 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 NTRK2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TrkB expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NTRK2 locus and enabling the study of TrkB-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TrkB pathway restoration in tumor cells with silenced or reduced NTRK2 expression.

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