BTRC Knockout Cell Line (Hela)

The choice depends on whether you are studying BTRC (β-TrCP1, FBXW1)'s role as a major SCF E3 ligase substrate receptor or modeling its broad biological functions. The Knockout line is the standard tool for asking whether β-TrCP is required for these processes — BTRC is one of two β-TrCP paralogs (BTRC/FBXW1 and FBXW11/β-TrCP2) that serve as F-box substrate receptors in SCF^β-TrCP E3 ubiquitin ligase complexes; β-TrCP recognizes phosphodegron motifs (DSGxxS) on diverse substrates including IκBα (NF-κB signaling), β-catenin (Wnt signaling, recognized by both β-TrCP and APC pathways), Cdc25A (DNA damage response), Emi1, REST, and many others. Overexpression is useful for studying BTRC in heterologous expression contexts. Important consideration: BTRC and FBXW11/β-TrCP2 share substantial substrate scope — single BTRC knockout may show modest phenotypes if FBXW11 compensates. For ubiquitin biology research, the EDITGENE BTRC Knockout in HeLa enables study of β-TrCP1-specific functions. Rescue with wild-type or substrate-binding-deficient BTRC is the standard specificity control. The knockout is valuable for studying NF-κB activation (IκBα degradation), Wnt signaling (β-catenin degradation), DNA damage response (Cdc25A degradation), and emerging targeted protein degradation approaches using β-TrCP-recruiting PROTACs.

Primary applications: • Substrate stability: IκBα, β-catenin, Cdc25A, Emi1, REST protein stability analysis (cycloheximide chase) in BTRC-null cells. • NF-κB signaling: TNF-α-induced IκBα degradation kinetics and downstream NF-κB activation. • Wnt signaling: β-catenin destruction complex analysis given β-TrCP's role. • FBXW11/β-TrCP2 paralog studies: FBXW11 expression analysis to interpret BTRC-specific functions. • β-TrCP-recruiting PROTAC platform development: emerging E3 ligase substrate receptor expansion beyond CRBN/VHL. EDITGENE recommends this model for researchers investigating β-TrCP-mediated ubiquitination, NF-κB regulation, and emerging β-TrCP-recruiting PROTACs.

Yes. BTRC rescue experiments require attention to F-box substrate receptor architecture: • Construct design: use a codon-modified BTRC sequence with a small C-terminal tag (FLAG, HA). BTRC has N-terminal F-box domain (SKP1 binding) and C-terminal WD40 domain (substrate phosphodegron binding) — preserve all elements. • Substrate-binding-deficient rescue: WD40 domain mutations disrupt phosphodegron substrate binding. • F-box-deficient rescue: F-box mutations disrupt SCF complex assembly. • Functional readout: rescue should restore IκBα/β-catenin/Cdc25A degradation. HeLa transduces efficiently with lentivirus and supports stable rescue line generation.