TFRC Knockout Cell Line (HEK293)

The choice depends on whether you are studying iron uptake biology, transferrin endocytosis, or using TFRC as a model receptor for clathrin-mediated endocytosis research. The Knockout line is the standard tool for asking whether TFRC is required for transferrin-mediated iron uptake — TFRC is the principal iron uptake receptor in most cell types and is essential for proliferating cells. Overexpression is useful for studying TFRC trafficking dynamics or for boosting iron uptake capacity. Important consideration: TFRC is essential in most proliferating cells — complete knockout may significantly impair viability and proliferation. The EDITGENE Knockout in HEK293 may exhibit reduced growth that should be characterized before extended use. Rescue with wild-type or endocytosis-deficient (cytoplasmic tail mutant) TFRC enables dissection of iron uptake from endocytic membrane trafficking functions. TFRC is also being explored as an antibody-drug conjugate target — the knockout serves as a critical specificity control for anti-TFRC therapeutics.

Primary applications: • Iron uptake assays: ⁵⁹Fe or fluorescent iron sensor measurements of transferrin-mediated iron uptake in the absence of TFRC. • Endocytosis studies: TFRC has long served as a model for clathrin-mediated endocytosis — fluorescent transferrin uptake kinetics characterize endocytic pathway integrity. • Anti-TFRC ADC validation: TFRC is a target for antibody-drug conjugates targeting proliferating cells; the knockout serves as a critical specificity control. • Viability and proliferation: TFRC is essential in most proliferating cells; characterization of growth phenotypes informs experimental design. EDITGENE recommends this model for researchers investigating iron homeostasis, clathrin-mediated endocytosis mechanisms, and TFRC-targeting therapeutics.

Yes. TFRC rescue experiments require attention to essentiality and endocytic function: • Construct design: use a codon-modified TFRC sequence with a C-terminal tag (FLAG, HA). TFRC is a type II membrane protein — N-terminal cytoplasmic tag is preferred over interfering with the extracellular transferrin-binding domain. • Viability considerations: TFRC is essential for proliferating cells; rescue experiments may need to be timed before significant proliferation defects accumulate. • Endocytosis-deficient rescue: cytoplasmic tail mutations (e.g., Y20A, internalization motif disruption) separate iron uptake from endocytic membrane trafficking functions. • Functional readout: rescue should restore transferrin binding (flow cytometry with fluorescent transferrin) and iron uptake capacity. HEK293 transduces efficiently with lentivirus, though rescue should be initiated promptly given TFRC's essential role in proliferation.