Functional CRISPR screening identifies the ufmylation pathway as a regulator of SQSTM1/p62

• DOI: 10.7554/elife.17290
• Publication Date: 2016-06-29T10:15:05Z
• Authors: Rowena DeJesus; Francesca Moretti; Gregory McAllister; Zuncai Wang; Phil Bergman; Shanming Liu; Elizabeth Frias; John Alford; John S Reece-Hoyes; Alicia Lindeman; Jennifer Kelliher; Carsten Russ; Judith Knehr; Walter Carbone; Martin Beibel; Guglielmo Roma; Aylwin Ng; John A Tallarico; Jeffery A Porter; Ramnik J Xavier; Craig Mickanin; Leon O Murphy; Gregory R Hoffman; Beat Nyfeler
• Keywords: Genetic screen; Cell fate determination

SQSTM1 is an adaptor protein that integrates multiple cellular signaling pathways and whose expression tightly regulated at the transcriptional post-translational level. Here, we describe a forward genetic screening paradigm exploiting CRISPR-mediated genome editing coupled to cell selection step by FACS identify regulators of SQSTM1. Through systematic comparison pooled libraries, show CRISPR superior RNAi in identifying known modulators. A genome-wide screen exposed MTOR signalling entire macroautophagy machinery as key identified several novel modulators including HNRNPM, SLC39A14, SRRD, PGK1 ufmylation cascade. We regulates eliciting type-specific ER stress response which induces results its accumulation cytosol. This study validates powerful method map repertoire regulate fate individual target protein.