ILDR1 CRISPR Activation Plasmid (m2): sc-430649-ACT-2

Mouse Ildr1 encodes immunoglobulin-like domain-containing receptor 1 (ILDR1), a single-pass transmembrane protein enriched at tricellular tight junctions where it helps organize epithelial barrier architecture and regulates paracellular permeability. ILDR1 participates in junctional assembly and signaling processes linked to apical–basal polarity and membrane protein trafficking, supporting coordinated cell–cell adhesion in sensory and epithelial tissues. Disruption of ILDR1 function is associated with tight-junction defects and auditory pathophysiology, making Ildr1 a relevant target for studying mechanisms of hearing loss and epithelial barrier dysfunction. Gene editing of mouse Ildr1 enables in vivo and in vitro interrogation of tricellular junction biology, tissue-specific barrier regulation, and downstream transcriptomic or proteomic changes in models of sensory epithelial development and disease-relevant stress responses.

ILDR1 CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Ildr1 expression without altering the underlying DNA sequence.

ILDR1 CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ildr1 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 Ildr1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ILDR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ildr1 locus and enabling the study of ILDR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ILDR1 pathway restoration in tumor cells with silenced or reduced Ildr1 expression.

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