UNC93B1 CRISPR Activation Plasmid (h): sc-404369-ACT

UNC93B1 (UNC93 homolog B1) is an endoplasmic reticulum membrane protein that controls trafficking and signaling competence of nucleic acid–sensing Toll-like receptors, including TLR3, TLR7, TLR8, and TLR9. By chaperoning these receptors from the ER to endolysosomal compartments, UNC93B1 helps set the threshold for type I interferon and NF-κB pathway activation downstream of pathogen- and damage-associated nucleic acids. Altered UNC93B1 function can perturb innate immune sensing, cytokine production, and antigen-presenting cell activation, linking it to immune dysregulation and susceptibility to infection-driven inflammatory phenotypes. As a regulator of endosomal TLR compartmentalization, UNC93B1 is frequently studied in monocytes, dendritic cells, B cells, and epithelial models to dissect innate immune circuitry.

UNC93B1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UNC93B1 expression without altering the underlying DNA sequence.

UNC93B1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UNC93B1 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 UNC93B1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UNC93B1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UNC93B1 locus and enabling the study of UNC93B1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UNC93B1 pathway restoration in tumor cells with silenced or reduced UNC93B1 expression.

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