CKAP4 Knockout HUH-6 Cell Line

The choice depends on whether you are studying CKAP4's role as a DKK1 (Dickkopf-1) receptor in cancer or modeling its functions in ER membrane biology. The Knockout line is the standard tool for asking whether CKAP4 is required for these processes — CKAP4 (cytoskeleton-associated protein 4, CLIMP-63, p63) was originally characterized as an ER membrane protein involved in ER morphology; recent breakthrough studies (Kimura et al. JCI 2016, Cancer Research 2019; Tsujiuchi-Kikawa-Sakamoto group) identified CKAP4 at the cell surface as a novel DKK1 receptor, distinct from the canonical LRP6-DKK1 pathway, driving PI3K-AKT signaling and cancer cell proliferation. Overexpression is useful for studying CKAP4 gain-of-function effects. For cancer and Wnt biology research, the EDITGENE CKAP4 Knockout in HuH-6 is highly relevant — HuH-6 is a hepatoblastoma cell line, and CKAP4 has been characterized as a poor prognostic marker in pancreatic, lung, and other cancers driving DKK1-AKT signaling. Rescue with wild-type or DKK1-binding-deficient CKAP4 enables structure-function studies. The knockout is a critical specificity tool for emerging anti-CKAP4 antibodies and CKAP4-DKK1 axis-targeted cancer therapeutics — CKAP4-DKK1 represents a novel therapeutic axis distinct from canonical Wnt-Frizzled-LRP pathway.

Primary applications: • DKK1-AKT signaling: DKK1-induced phospho-AKT (S473, T308) analysis in CKAP4-null cells — CKAP4 mediates non-canonical DKK1 oncogenic signaling. • Hepatoblastoma biology: in HuH-6 context, CKAP4-DKK1 axis-driven proliferation and survival assays. • ER membrane biology: ER tubule/sheet morphology analysis given CKAP4's ER membrane role. • Anti-CKAP4 therapy development: critical specificity control for emerging anti-CKAP4 antibodies and CKAP4-DKK1 axis-targeted cancer therapeutics. EDITGENE recommends this hepatoblastoma model for researchers investigating CKAP4-DKK1 oncogenic axis and emerging novel Wnt-pathway-adjacent therapeutic strategies.

Yes. CKAP4 rescue experiments require attention to dual localization: • Construct design: use a codon-modified CKAP4 sequence with a small intracellular C-terminal tag (FLAG, HA). CKAP4 is a type II membrane protein with single transmembrane span; both ER-resident (palmitoylated) and surface (DKK1 receptor) pools exist — preserve membrane topology. • Surface localization validation: confirm cell surface CKAP4 by surface biotinylation or FACS before DKK1 binding studies. • Palmitoylation-deficient rescue: C100A palmitoylation site mutations may alter ER/surface distribution. • DKK1-binding-deficient rescue: extracellular domain mutations disrupt DKK1 binding. • Functional readout: rescue should restore DKK1-induced phospho-AKT signaling. HuH-6-specific considerations: • HuH-6 is a human hepatoblastoma cell line — a pediatric liver tumor model relevant for hepatoblastoma biology and Wnt-driven cancer studies (HuH-6 has CTNNB1 mutation). • Lentiviral transduction is supported with moderate efficiency. • HuH-6 retains some hepatocyte features and is one of the principal hepatoblastoma cell line models distinct from HCC lines (Hep-G2, Huh-7).