Iglon5 CRISPR Activation Plasmid (m): sc-431673-ACT

Iglon5 (IgLON family member 5) is a glycosylphosphatidylinositol (GPI)-anchored cell surface adhesion molecule predominantly associated with neuronal membranes, where it contributes to neurite outgrowth, synaptic organization, and cell–cell recognition. As part of the IgLON immunoglobulin superfamily, Iglon5 participates in processes linked to axon guidance and neuronal network maturation, interfacing with membrane microdomains and extracellular interactions that shape circuit connectivity. Dysregulation of IG/LON family adhesion programs has been connected to altered synaptic stability and neuroinflammatory signaling contexts, supporting research interest in neurodegeneration and sleep-associated neurological phenotypes. In mouse systems, Iglon5 is commonly studied in relation to neuronal development, synaptic maintenance, and stress-responsive changes in cell-surface proteomes.

IGLON5 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Iglon5 expression without altering the underlying DNA sequence.

IGLON5 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Iglon5 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 Iglon5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IGLON5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Iglon5 locus and enabling the study of IGLON5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IGLON5 pathway restoration in tumor cells with silenced or reduced Iglon5 expression.

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