UNC5H2 CRISPR Activation Plasmid (h): sc-402347-ACT

UNC5B encodes the netrin receptor UNC5H2, a transmembrane guidance cue receptor that regulates axon navigation, neuronal migration, and cell survival through netrin-dependent signaling and dependence receptor behavior in the absence of ligand. UNC5H2 modulates cytoskeletal dynamics and adhesion via Rho-family GTPases and focal adhesion-associated pathways, influencing directional motility and tissue patterning during development. In non-neuronal contexts, UNC5B/UNC5H2 contributes to vascular and epithelial organization and can affect apoptosis, proliferation, and invasion programs. Dysregulated UNC5B signaling has been associated with altered cell migration and survival phenotypes relevant to neurodevelopmental disorders and cancer biology, making it a useful target for mechanistic pathway studies.

UNC5H2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UNC5B expression without altering the underlying DNA sequence.

UNC5H2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UNC5B 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 UNC5B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UNC5H2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UNC5B locus and enabling the study of UNC5H2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UNC5H2 pathway restoration in tumor cells with silenced or reduced UNC5B expression.

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