HEK293-A mRFP-ATG9 Cell Line

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.

Contributors

Inventor Institute
Sharon Tooze Cancer Research UK, London Research Institute: Lincoln's Inn Fields
Cat. #: 151828
Tool sub type: Continuous
Unit size: 1×10^6 cells / vial
Research Fields: Apoptosis and autophagy;Cancer;Cell biology;Cell signaling and signal transduction;Drug development;Metabolism
Organism: Human
Tissue: Embryo
Model: Reporter
Product 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.
Conditional: Yes
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.
Additional notes: HEK293 cells were obtained under licence from AdVec
Cellosaurus ID: CVCL_W349
Parental cell line: HEK293-A
Format: Frozen
Storage conditions: Liquid Nitrogen
Shipping conditions: Dry ice
Growth medium: DMEM + 10% FCS + 2mM Glutamine + 400ug/ml geneticin (G418) for maintainance.
Recommended controls: HEK293-A parental line
Mycoplasma free: Yes
Biosafety level: 1
References:

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.

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