PT  - JOURNAL ARTICLE
AU  - Christopher J Shoemaker
AU  - Tina Q Huang
AU  - Nicholas R Weir
AU  - Nicole Polyakov
AU  - Vladimir Denic
TI  - A CRISPR screening approach for identifying novel autophagy-related factors and cytoplasm-to-lysosome trafficking routes
AID  - 10.1101/229732
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 229732
4099  - http://biorxiv.org/content/early/2017/12/22/229732.short
4100  - http://biorxiv.org/content/early/2017/12/22/229732.full
AB  - Selective autophagy comprises cytoplasm-to-lysosome trafficking routes that transport cargos using double-membrane vesicles (autophagosomes). Cargos are detected by receptor proteins, which typically also bind to lipid-conjugated LC3 proteins on autophagosome membranes. We dissected lysosomal delivery of four SQSTM1-like receptors by genome-wide CRISPR screening looking for novel autophagy-related (ATG) factors and trafficking routes. We uncovered new mammalian ATG factors including TMEM41B, an endoplasmic reticulum membrane protein required for autophagosome membrane expansion and/or closure. Furthermore, we found that certain receptors remain robustly targeted to the lysosome even in the absence of ATG7 or other LC3 conjugation factors. Lastly, we identified a unique genetic fingerprint behind receptor flux in ATG7KO cells, which includes factors implicated in nucleating autophagosome formation and vesicle trafficking factors. Our work uncovers new ATG factors, reveals a malleable network of autophagy receptor genetic interactions, and provides a valuable resource (http://crispr.deniclab.com) for further mining of novel autophagy mechanisms.