LAR CRISPR Activation Plasmid (m): sc-422521-ACT

Mouse Ptprf encodes the receptor-type protein tyrosine phosphatase LAR (PTPRF), a transmembrane signaling regulator that balances phosphorylation-dependent pathways controlling cell adhesion, migration, and neurite outgrowth. LAR modulates focal adhesion and cytoskeletal dynamics through interactions with extracellular matrix cues and intracellular scaffolds, influencing integrin-linked signaling and growth factor receptor outputs. In neuronal and metabolic tissues, PTPRF/LAR has been connected to synapse organization, axon guidance, and regulation of insulin receptor signaling, making it relevant to studies of neurodevelopmental phenotypes and insulin sensitivity mechanisms. Dysregulated phosphatase activity and altered adhesion signaling have also been implicated in oncogenic processes such as aberrant proliferation and invasion, supporting its use as a node for pathway interrogation.

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

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

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