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HEK293-A mRFP-ATG9

HEK293-A mRFP-ATG9

Catalogue No.

17100531

Cell Line Name

HEK293-A mRFP-ATG9

Cell Line Description

The HEK293-A mRFP-ATG9 cell line is a stable cell line expressing mRFP-tagged ATG9 (human) in background of HEK293A cell line.

General Info

Species

Human

Unique to ECACC

Yes

Release Conditions

Restricted - commercial organisations are required to complete the 'Cell Line Release Authorisation for Research Use in Commercial Organisations' release conditions form in the supporting documents section.

Characteristics

Tissue of Origin

Embryonic Kidney

Morphology

Neuronal

DNA profile (STR Profile)

Amelogenin: X
CSF1PO: 12
D3S1358: 15,17
D5S818: 8
D7S820: 11
D8S1179: 12,14
D13S317: 12,14
D16S539: 9,13
D18S51: 17
D21S11: 30.2
FGA: 21,23
Penta D: 9
Penta E: 7,15
TH01: 7,9.3
TPOX: 11
vWA: 16,19

Applications

HEK293-A mRFP-ATG9 can be used to monitor delivery of membrane from the endosomes and golgi to the forming autophagosome using live-cell microscopy and biochemical approaches.

Disease

Autophagy

Culture Conditions

Cell Type

Neuronal

Subculture Routine

Trypsinise cultures at 80% confluence with 0.05% Trypsin/EDTA. Seed flasks at 2x10⁴ cells/ cm². Cultures must be incubated in a humidified 10% CO₂/90% air incubator at 37°C.

Culture Medium

DMEM + 10% FBS + 2mM Glutamine + 400 µg/ml geneticin (G418) for maintenance.

Growth Mode

Adherent

Additional Info

Depositor

Ximbio (formerly known as Cancer Research Technologies Limited)

Country of Origin

United Kingdom

GMO Status

Genetically Modified Organism Class 1 (GMO1)

Genetic Modification

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 800µg/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.

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

Orsi A, Razi M, Dooley HC, Robinson D, Weston AE, Collinson LM, Tooze SA. Dynamic and transient interactions of Atg9 with autophagosomes, but not membrane integration, are required for autophagy.Mol Biol Cell. 2012 May;23(10):1860-73. doi: 10.1091/mbc. E11-09-0746. Epub 2012 Mar 28. DOI: 10.1091/mbc . E11-09-0746. PMCID: PMC3350551. PMID: 22456507 [Indexed for MEDLINE]. Young AR, Chan EY, Hu XW, Köchl R, Crawshaw SG, High S, Hailey DW, Lippincott-Schwartz J, Tooze SA. Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes. J Cell Sci. 2006 Sep 15;119(Pt 18):3888-900. Epub 2006 Aug 29. DOI: 10.1242/jcs.03172 PMID: 16940348 [Indexed for MEDLINE]

Available Formats

  • Frozen

If use of this culture results in a scientific publication, it should be cited in the publication as: HEK293-A mRFP-ATG9 (ECACC 17100531).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.