KLRG1 CRISPR Activation Plasmid (m): sc-424537-ACT

Mouse Klrg1 encodes KLRG1, an inhibitory C-type lectin-like receptor prominently expressed on differentiated effector and memory subsets of NK cells and CD8+ T cells. By engaging classical cadherins such as E-cadherin on target cells, KLRG1 transduces inhibitory signals that modulate cytotoxicity, cytokine output, and proliferative capacity, linking adhesion cues to immune checkpoint-like regulation. KLRG1 is widely used as a marker of immune cell maturation and functional state, and altered KLRG1 signaling has been associated with changes in antiviral and antitumor immune responses, immune aging, and chronic inflammation. These features make Klrg1 a relevant node for studying immune regulation, tissue surveillance, and effector cell persistence in disease models.

KLRG1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Klrg1 expression without altering the underlying DNA sequence.

KLRG1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Klrg1 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 Klrg1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KLRG1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Klrg1 locus and enabling the study of KLRG1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KLRG1 pathway restoration in tumor cells with silenced or reduced Klrg1 expression.

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