BTN2A2 CRISPR Activation Plasmid (m): sc-433634-ACT

Mouse Btn2a2 encodes BTN2A2, a butyrophilin family immunoregulatory surface protein that modulates T cell activation and peripheral tolerance through contact-dependent signaling with antigen-presenting cells. BTN2A2 activity influences immune synapse function, cytokine production, and lymphocyte proliferation, linking it to pathways that shape adaptive immune responses in lymphoid tissues. Altered BTN2A2 expression has been studied in contexts of chronic inflammation and autoimmune-like phenotypes, where shifts in co-regulatory signaling can affect effector versus regulatory T cell balance. As a checkpoint-associated molecule, BTN2A2 is also relevant to tumor immunology research focused on mechanisms of immune evasion and T cell dysfunction.

BTN2A2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Btn2a2 expression without altering the underlying DNA sequence.

BTN2A2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Btn2a2 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 Btn2a2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BTN2A2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Btn2a2 locus and enabling the study of BTN2A2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BTN2A2 pathway restoration in tumor cells with silenced or reduced Btn2a2 expression.

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