PDCD1LG2/PD-L2 CRISPR Activation Plasmid (m): sc-425483-ACT

Pdcd1lg2 encodes PDCD1LG2/PD-L2, an immune checkpoint ligand expressed on antigen-presenting cells and inducible on some stromal and tumor-associated cell types that engages PD-1 on activated T cells. PD-L2–PD-1 signaling dampens TCR-driven activation, cytokine production, and proliferation through inhibitory phosphatase recruitment and downstream attenuation of PI3K–AKT and MAPK pathways, shaping peripheral tolerance and inflammatory set points. In mouse models, Pdcd1lg2 regulation contributes to control of T cell exhaustion, Th1/Th2 polarization, and local immune privilege within tissues. Dysregulated PD-L2 expression is studied in contexts including chronic infection, autoimmunity, transplant immunobiology, and tumor microenvironment immune evasion as a mechanistic biomarker of immune suppression.

PDCD1LG2/PD-L2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Pdcd1lg2 expression without altering the underlying DNA sequence.

PDCD1LG2/PD-L2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Pdcd1lg2 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 Pdcd1lg2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PDCD1LG2/PD-L2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Pdcd1lg2 locus and enabling the study of PDCD1LG2/PD-L2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PDCD1LG2/PD-L2 pathway restoration in tumor cells with silenced or reduced Pdcd1lg2 expression.

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