FOXP3 CRISPR Activation Plasmid (m): sc-422896-ACT

Mouse Foxp3 encodes FOXP3, a forkhead/winged-helix transcription factor that serves as a lineage-defining regulator of regulatory T cells and a central controller of immune tolerance. FOXP3 coordinates transcriptional programs governing T cell receptor signaling thresholds, IL-2 pathway suppression, and chromatin-dependent control of cytokine gene networks, thereby shaping peripheral immune homeostasis. Disruption or dysregulated expression of FOXP3 perturbs Treg differentiation and suppressive function, linking this axis to inflammatory and autoimmune-like phenotypes and altered responses in immune-mediated disease models. FOXP3 activity is also studied in the context of immune cell plasticity, tissue inflammation, and tumor-immune interactions where Treg programs influence local immune environments.

FOXP3 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Foxp3 expression without altering the underlying DNA sequence.

FOXP3 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Foxp3 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 Foxp3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FOXP3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Foxp3 locus and enabling the study of FOXP3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FOXP3 pathway restoration in tumor cells with silenced or reduced Foxp3 expression.

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