TAB2 Knockout HAP1 Cell Line

The choice depends on whether you are studying TAB2's role as a TAK1 adapter in IL-1/TNF/TLR signaling, or its functions in NF-κB pathway activation. The Knockout line is the standard tool for both — TAB2 binds K63-linked polyubiquitin chains and recruits TAK1 to signaling complexes, making it essential for cytokine-induced NF-κB activation. Overexpression is useful for testing TAB2's sufficiency in pathway activation or for studying its competition with TAB3 paralog. For innate immune signaling research, the EDITGENE TAB2 Knockout in HAP1 is informative for dissecting K63-ubiquitin-dependent signaling, though TAB3 paralog compensation should be considered when interpreting phenotypes. Rescue with wild-type or ubiquitin-binding-deficient (NZF domain mutant) TAB2 separates ubiquitin-sensing from adapter functions.

Primary applications: • NF-κB activation: NF-κB reporter assays and target gene expression following IL-1β, TNF-α, or TLR stimulation to assess TAB2's adapter function. • TAK1 complex assembly: co-immunoprecipitation analysis of TAK1-TAB1-TAB2 complex formation following K63-ubiquitin signal generation. • Cytokine signaling: phospho-IKK, IκBα degradation, and downstream cytokine gene induction following innate immune stimulation. • Paralog studies: TAB3 expression measurement to interpret TAB2-specific phenotypes given their overlapping function in TAK1 complex assembly. EDITGENE recommends this model for researchers investigating innate immune signaling, TAK1 pathway biology, and NF-κB activation mechanisms.

Yes. TAB2 rescue experiments require attention to its ubiquitin-binding and adapter functions: • Construct design: use a codon-modified TAB2 sequence with a small C-terminal tag (FLAG, HA). The C-terminal NZF (nuclear protein localization 4-type zinc finger) domain mediates K63-ubiquitin chain binding and must be preserved. • NZF mutant rescue: specific NZF domain mutations (e.g., T674A) abolish K63-polyubiquitin binding without disrupting TAK1 interaction — the gold standard for separating ubiquitin-sensing from adapter functions. • Paralog considerations: TAB3 expression analysis helps interpret single TAB2 rescue, given their overlapping roles in TAK1 complex assembly. • Functional readout: rescue should restore IL-1β-, TNF-α-, and TLR-induced NF-κB activation and TAK1 phosphorylation patterns. HAP1-specific considerations: • Diploidization: HAP1 cells gradually diploidize during extended culture — confirm ploidy by flow cytometry at the time of phenotypic assay. • Integration site sensitivity: position effects on transgene expression are more pronounced in near-haploid backgrounds; generating multiple independent rescue clones is strongly recommended. • Transduction efficiency: HAP1 transduces with lentivirus at moderate efficiency — increase MOI compared to standard immortalized lines.