Lingo-2 CRISPR Activation Plasmid (h): sc-414778-ACT

Human LINGO2 encodes Lingo-2, a single-pass transmembrane leucine-rich repeat and Ig-like domain protein implicated in regulating cell–cell signaling at the plasma membrane. Lingo-2 has been studied in the context of neural development and synaptic processes, where LINGO family proteins can influence axon guidance, neurite outgrowth, and receptor complex assembly. Altered LINGO2 expression has been associated in genetic and transcriptomic studies with neurological traits and disorders, supporting its relevance for investigating signaling programs that shape neuronal connectivity and cellular differentiation. As a membrane-associated modulator, Lingo-2 provides a tractable entry point for probing pathway-level changes through downstream transcriptional responses and phospho-signaling readouts.

Lingo-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LINGO2 expression without altering the underlying DNA sequence.

Lingo-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LINGO2 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 LINGO2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Lingo-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LINGO2 locus and enabling the study of Lingo-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Lingo-2 pathway restoration in tumor cells with silenced or reduced LINGO2 expression.

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