- Name Sharon Tooze
- Institute Cancer Research UK, London Research Institute: Lincoln's Inn Fields
- Tool name: HEK293-A mRFP-ATG9 Cell Line
- Tool type: Cell Lines
- Tool sub-type: Continuous
- Parental cell line: HEK293-A
- Organism: Human
- Tissue: Embryo
- Model: Reporter
- Conditional: Yes
- Description: The HEK293-A mRFP-ATG9 cell line is a stable cell line expressing mRFP-tagged ATG9 (human) in background of HEK293A cell line. It can be used to monitor delivery of membrane from the endosomes and Golgi to the forming autophagosome using live-cell microscopy and biochemical approaches.
- Research area: Cancer; Apoptosis and Programmed Cell Death; Cell Signaling & Signal Transduction; Cellular Transport; Drug Discovery & Development; Metabolism
- Production details: pEGFP-mRFP-Atg9 plasmid was transfected into HEK293A cells. The plasmid was generated by excision of the EGFP coding sequence from pEGFP-C1 and insertion of the mRFP-ATG9 cassette. The cells were selected with 800ug/ml geneticin (G418) followed by single cell clonal dilution. Clones were screened by confocal microscopy for low-level expression in which mRFP- Atg9 distribution resembled that of endogenous mAtg9 and by western blot. Clone 1F10 was selected as the best based on these parameters.
- Cellosaurus ID: CVCL_W349
- Additional notes: HEK293 cells were obtained under licence from AdVec
- For Research Use Only
- Target: ATG9A
- Application notes: HEK293 cells were obtained under licence from AdVec
- Format: Frozen
- Growth medium: DMEM + 10% FCS + 2mM Glutamine + 400ug/ml geneticin (G418) for maintainance.
- Storage conditions: Liquid Nitrogen
- Shipping conditions: Dry ice
- Mycoplasma free: Yes
- Biosafety level: 1
- • Orsi et al. 2012. Mol Biol Cell. 23(10):1860-73. PMID: 22456507.
- • Dynamic and transient interactions of Atg9 with autophagosomes, but not membrane integration, are required for autophagy.
- • Young et al. 2006. J Cell Sci. 119(Pt 18):3888-900. PMID: 16940348.
- • Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes.