I am looking to create a set of mammalian cell line (CHOk1) witch are deficient in the autophagy response. I need to find two genes to knockout in my cells to prevent autophagy (currently i am targeting ATG5 but i am not sure what other gen to target alongside this)?
Also I would like to try and make a cell line which is able undergo autopohagy but is unable to fuse the autophagosome with the lysosomes (blocking of autophagy flux) via gene knockout. What gene(s) would be best to knockout to achieve this?
Check this guide Article Guidelines for the use and interpretation of assays for moni...
not an easy question, though you can follow this link and decide yourself: http://clincancerres.aacrjournals.org/content/21/3/498
alternatively, you can chemically induce or repress autophagy. a list of chemical compounds that might be useful is here https://www.selleckchem.com/autophagy.html
good luck!
ULK1 regulates autophagy as an upstream molecule, so that why not knockdown ULK1 with Atg5 and/or Atg7?
Liam Davison-Gates To block autophagy flux you can knock down Rab7, syntaxin17, VPS33A, VPS16, kinesin KIF5B
For blocking autophagosome-lysosome fusion, you can target Syntaxin17. See RNAi/ko phenotypes in this paper: Article The Hairpin-type Tail-Anchored SNARE Syntaxin 17 Targets to ...
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