A20/TNFAIP3 Lentiviral Activation Particles (m): sc-423436-LAC

Tnfaip3 encodes A20/TNFAIP3, a ubiquitin-editing enzyme that terminates inflammatory signaling by restricting NF-κB and MAPK pathway activation downstream of TNF receptor, Toll-like receptor, and IL-1 receptor inputs. Through coordinated deubiquitination and ubiquitin ligase activities, A20 modulates K63- and M1-linked ubiquitin scaffolds on key adapters such as RIPK1, TRAF6, and NEMO to control cytokine production and cell-death signaling. In mouse systems, altered A20 activity perturbs innate and adaptive immune homeostasis, influencing responses in myeloid and lymphoid compartments and shaping tissue inflammation. These functions make Tnfaip3 a common node studied in inflammation biology, autoimmunity-associated mechanisms, and oncogenic signaling contexts where NF-κB regulation is central.

A20/TNFAIP3 Lentiviral Activation Particles (m) address this need by packaging the complete synergistic activation mediator (SAM) transcriptional activation system into transduction-ready, high-titer lentiviral particles, enabling efficient Tnfaip3 upregulation across a broader range of human cell types.

A20/TNFAIP3 Lentiviral Activation Particles (m) deliver all functional components of the synergistic activation mediator (SAM) system via lentiviral transduction. The system comprises three particle preparations co-transduced into target cells: one encoding catalytically inactive dCas9 (D10A and N863A mutations) fused to the VP64 transactivation domain with a blasticidin resistance gene; one encoding the MS2-p65-HSF1 fusion protein with a hygromycin resistance gene; and one encoding a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers with a puromycin resistance gene. Following lentiviral transduction and genomic integration of the expression cassettes, the SAM components are stably expressed and assemble at the target locus within the proximal promoter region upstream of the Tnfaip3 transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous A20/TNFAIP3 expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native Tnfaip3 genomic locus and regulatory architecture.

The lentiviral format offers several practical advantages: stable genomic integration supports heritable activation across cell divisions; high-titer particle preparations eliminate the need for in-house viral production; and compatibility with primary, non-dividing, and transfection-resistant cell types expands experimental accessibility. Successful transduction can be confirmed and enriched through triple antibiotic selection using puromycin, hygromycin, and blasticidin.

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