TFE3 Knockout Cell Line (HEK293)

The choice depends on whether you are studying TFE3's role in lysosomal biogenesis (CLEAR network), autophagy, or modeling TFE3 fusion oncoproteins in cancer. The Knockout line is appropriate for asking whether TFE3 is required for these processes — TFE3 is part of the MiT/TFE family (with TFEB, MITF, TFEC) and shares substantial functional overlap. Overexpression is useful for studying constitutive lysosomal/autophagy gene activation or for modeling TFE3 fusions seen in translocation renal cell carcinoma and alveolar soft part sarcoma. For TFE3 research, an important consideration is the strong functional redundancy with TFEB — single TFE3 knockout often shows mild basal phenotypes because TFEB compensates. The EDITGENE Knockout in HEK293 is most informative when combined with TFEB status assessment. Rescue with wild-type, phosphomimetic (constitutively cytoplasmic), or phospho-deficient (constitutively nuclear) TFE3 enables dissection of mTORC1-regulated nuclear translocation.

Primary applications: • CLEAR network gene expression: analysis of lysosomal biogenesis and autophagy gene expression (LAMP1, CTSD, ATG9, GABARAPL1) in the absence of TFE3. • Subcellular localization studies: nuclear/cytoplasmic distribution of TFE3 family members under nutrient stress, lysosomal stress, and mTORC1 inhibition. • MiT/TFE redundancy studies: TFEB and TFE3 share substantial overlap — combined analysis with TFEB knockdown or knockout reveals shared versus TFE3-specific functions. • TFE3 fusion oncoprotein studies: rescue with wild-type or fusion-mutant TFE3 (mimicking translocation RCC variants) enables cancer biology research. EDITGENE recommends this model for researchers investigating lysosomal biogenesis, autophagy regulation, and TFE3 fusion-driven cancers.

Yes. TFE3 rescue experiments require attention to its mTORC1-regulated localization: • Construct design: use a codon-modified TFE3 sequence with a small C-terminal tag (FLAG, HA). TFE3 contains an N-terminal transactivation domain — large N-terminal tags should be avoided. • Localization-variant rescue: phosphomimetic (S321E, constitutively cytoplasmic) and phospho-deficient (S321A, constitutively nuclear) variants enable dissection of mTORC1-regulated nuclear translocation. • Fusion oncoprotein rescue: TFE3 fusion proteins (PRCC-TFE3, ASPSCR1-TFE3) can be introduced to model translocation RCC and alveolar soft part sarcoma. • Functional readout: rescue should restore CLEAR network gene expression (LAMP1, CTSD, TFEB, GABARAPL1) and TFE3 nuclear translocation following mTORC1 inhibition. HEK293 transduces efficiently with lentivirus and supports TFE3 rescue with high reproducibility for transcription factor research applications.