Latrophilin-2 CRISPR Activation Plasmid (m): sc-430273-ACT

Adgrl2 encodes latrophilin-2, an adhesion G protein–coupled receptor (aGPCR) that contributes to cell–cell communication by coupling extracellular adhesion cues to intracellular signaling. Latrophilin-2 participates in processes linked to neuronal connectivity, synapse organization, and developmental patterning through GPCR-associated signaling and cytoskeletal remodeling. By engaging extracellular ligands and adhesion modules, it can influence pathways that regulate cell migration, polarity, and tissue architecture. Altered aGPCR signaling and adhesion programs are frequently studied in the context of neurodevelopmental phenotypes and dysregulated cell interactions relevant to disease-associated circuit function.

Latrophilin-2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Adgrl2 expression without altering the underlying DNA sequence.

Latrophilin-2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Adgrl2 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 Adgrl2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Latrophilin-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Adgrl2 locus and enabling the study of Latrophilin-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Latrophilin-2 pathway restoration in tumor cells with silenced or reduced Adgrl2 expression.

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