FBXW7 Knockout Cell Line (Hela)

The choice depends on whether you are studying FBXW7's role as the principal F-box substrate receptor of the SCF^FBXW7 E3 ubiquitin ligase or modeling FBXW7-mutation-driven cancers. The Knockout line is the standard tool for asking whether FBXW7 is required for these processes — FBXW7 (FBW7, CDC4) is a tumor suppressor F-box protein that recognizes phosphodegron motifs (CPD, Cdc4 phosphodegron, T/S-P-X-X-S/T/E) on diverse substrates including MYC, Cyclin E, Notch1 ICD, Jun, mTOR, MCL1, SREBP1; FBXW7 is one of the most frequently mutated tumor suppressors in cancer (T-ALL, colorectal cancer, endometrial cancer, cholangiocarcinoma). Overexpression is useful for studying FBXW7 in heterologous expression contexts. For cancer biology research, the EDITGENE FBXW7 Knockout in HeLa is highly informative — FBXW7 loss stabilizes its substrates (MYC, Cyclin E, Notch1), driving proliferation and transformation. Rescue with wild-type or substrate-binding-deficient FBXW7 (e.g., R465H, R479Q, R505C hot-spot mutations in the WD40 substrate-binding domain) enables disease genotype-function studies of common cancer mutations. The knockout is valuable for studying substrate stability of MYC, Cyclin E, Notch1, mTOR pathway proteins, and emerging FBXW7-restored cancer therapeutic strategies.

Primary applications: • Substrate protein stability: MYC, Cyclin E, Notch1 (intracellular domain), Jun, mTOR, MCL1, SREBP1 protein stability and half-life analysis (cycloheximide chase) in FBXW7-null cells. • Cancer mutation modeling: rescue with hot-spot mutations (R465H, R479Q, R505C in the WD40 substrate-binding domain) for genotype-function studies of common FBXW7 cancer mutations. • T-ALL biology: in heterologous T-ALL-relevant contexts, characterization of FBXW7-NOTCH1 axis in T-cell acute lymphoblastic leukemia. • Substrate phosphodegron analysis: assessment of CPD-dependent substrate recognition given FBXW7's requirement for substrate phosphorylation. EDITGENE recommends this model for researchers investigating FBXW7 tumor suppressor biology, substrate ubiquitination, and FBXW7-mutation-driven cancers.

Yes. FBXW7 rescue experiments are well-established for cancer ubiquitination research: • Construct design: use a codon-modified FBXW7 sequence with a small C-terminal tag (FLAG, HA). FBXW7 has N-terminal F-box (SKP1 binding) and C-terminal WD40 substrate-binding domain — preserve all elements. Three FBXW7 isoforms (α nuclear, β cytoplasmic, γ nucleolar) exist with distinct localizations. • Cancer mutation rescue: hot-spot mutations R465H, R479Q, R505C in the WD40 domain disrupt substrate phosphodegron binding — invaluable for studying common FBXW7 cancer mutations. • Substrate-binding-deficient rescue: WD40 domain mutations abolish substrate recognition without affecting SCF complex assembly. • Functional readout: rescue should restore MYC, Cyclin E, Notch1 degradation and reduce proliferative signaling. HeLa transduces efficiently with lentivirus and supports stable rescue line generation.