AMIGO2 CRISPR Activation Plasmid (h): sc-403312-ACT

AMIGO2 (adhesion molecule with Ig-like domain 2) is a type I transmembrane cell-surface protein in the AMIGO family that supports cell–cell interactions, neurite outgrowth, and axon guidance during nervous system development. In human cells, AMIGO2 has been linked to regulation of adhesion-dependent signaling, migration, and survival programs that intersect with cytoskeletal remodeling and receptor-mediated pathways. Altered AMIGO2 expression has been reported in contexts of tumor progression and metastasis-associated phenotypes, where changes in adhesion and motility can remodel tissue architecture. These features make AMIGO2 a useful target for studying how extracellular cues are coupled to intracellular signaling and differentiation states.

AMIGO2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AMIGO2 expression without altering the underlying DNA sequence.

AMIGO2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AMIGO2 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 AMIGO2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AMIGO2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AMIGO2 locus and enabling the study of AMIGO2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AMIGO2 pathway restoration in tumor cells with silenced or reduced AMIGO2 expression.

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