A20/TNFAIP3 Lentiviral Activation Particles (h): sc-400447-LAC

TNFAIP3 encodes A20, a zinc finger ubiquitin-editing enzyme that functions as a central negative regulator of inflammatory signaling. A20 terminates NF-κB activation downstream of TNF receptor, Toll-like receptors, and IL-1 receptor pathways by modulating K63- and M1-linked ubiquitin chains and promoting proteasomal turnover of key adaptor proteins. Through these actions, A20 constrains cytokine production, cell survival signaling, and innate immune activation, helping maintain tissue homeostasis. Dysregulated TNFAIP3 expression or function is associated with aberrant immune responses and has been implicated in autoimmune and inflammatory disease mechanisms, as well as signaling dependencies in certain malignancies.

A20/TNFAIP3 Lentiviral Activation Particles (h) 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 (h) 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.