LAR CRISPR Activation Plasmid (h): sc-402674-ACT

Human PTPRF encodes leukocyte common antigen-related (LAR), a receptor-type protein tyrosine phosphatase that modulates signal transduction by dephosphorylating key substrates at the plasma membrane. LAR regulates cell adhesion and migration through crosstalk with integrin and focal adhesion pathways, shaping cytoskeletal remodeling and downstream kinase signaling. It also influences receptor tyrosine kinase networks, including insulin receptor signaling, with effects on cellular growth and metabolic responses. Dysregulated PTPRF/LAR activity has been associated with altered adhesion-dependent signaling and has been studied in contexts such as cancer cell invasiveness and metabolic disease-relevant insulin sensitivity phenotypes.

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

LAR CRISPR Activation Plasmid (h) 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.