Abin-1 CRISPR Activation Plasmid (m): sc-425441-ACT

Mouse Tnip1 encodes ABIN-1 (A20-binding inhibitor of NF-κB), an ubiquitin-binding adaptor that constrains inflammatory signaling downstream of TNF receptor, Toll-like receptors, and IL-1 receptor pathways. ABIN-1 cooperates with the deubiquitinase A20/TNFAIP3 to modulate K63- and linear (M1) ubiquitin signaling on complexes containing RIPK1 and NEMO/IKK, thereby shaping NF-κB and MAPK transcriptional outputs. Through these interactions, ABIN-1 influences innate immune activation, cytokine production, and survival pathways, making it relevant to models of autoimmunity, inflammatory disease, and immune-driven tissue pathology. In addition, ABIN-1 has been linked to regulation of cell stress responses and can affect proliferative phenotypes in contexts where NF-κB signaling is dysregulated.

Abin-1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Tnip1 expression without altering the underlying DNA sequence.

Abin-1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Tnip1 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 Tnip1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Abin-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Tnip1 locus and enabling the study of Abin-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Abin-1 pathway restoration in tumor cells with silenced or reduced Tnip1 expression.

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