title: Genome-Wide CRISPR-Cas9 Screen Identifies MicroRNAs That Regulate Myeloid Leukemia Cell Growth.
abstract: Mammalian microRNA expression is dysregulated in human cancer. However, the functional relevance of many microRNAs in the context of tumor biology remains unclear. Using CRISPR-Cas9 technology, we performed a global loss-of-function screen to simultaneously test the functions of individual microRNAs and protein-coding genes during the growth of a myeloid leukemia cell line. This approach identified evolutionarily conserved human microRNAs that suppress or promote cell growth, revealing that microRNAs are extensively integrated into the molecular networks that control tumor cell physiology. miR-155 was identified as a top microRNA candidate promoting cellular fitness, which we confirmed with two distinct miR-155-targeting CRISPR-Cas9 lentiviral constructs. Further, we performed anti-correlation functional profiling to predict relevant microRNA-tumor suppressor gene or microRNA-oncogene interactions in these cells. This analysis identified miR-150 targeting of p53, a connection that was experimentally validated. Taken together, our study describes a powerful genetic approach by which the function of individual microRNAs can be assessed on a global level, and its use will rapidly advance our understanding of how microRNAs contribute to human disease.
authors.affiliation: Department of Pathology, University of Utah, Salt Lake City, Utah, United States of America.
authors.forename: Jared
authors.initials: J
authors.lastname: Wallace
authors.affiliation: Department of Pathology, University of Utah, Salt Lake City, Utah, United States of America.
authors.forename: Ruozhen
authors.initials: R
authors.lastname: Hu
authors.affiliation: Huntsman Cancer Institute, University of Utah, Salt Lake City, Utah, United States of America.
authors.forename: Timothy L
authors.initials: TL
authors.lastname: Mosbruger
authors.affiliation: Mutation Generation and Detection Core, HSC Core Research Facilities, University of Utah, Salt Lake City, Utah, United States of America.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Dahlem
authors.affiliation: Department of Pathology, University of Utah, Salt Lake City, Utah, United States of America.
authors.forename: W Zac
authors.initials: WZ
authors.lastname: Stephens
authors.affiliation: Department of Pathology, UCLA, Los Angeles, California, United States of America.
authors.forename: Dinesh S
authors.initials: DS
authors.lastname: Rao
authors.affiliation: Department of Pathology, University of Utah, Salt Lake City, Utah, United States of America.
authors.forename: June L
authors.initials: JL
authors.lastname: Round
authors.affiliation: Department of Pathology, University of Utah, Salt Lake City, Utah, United States of America.
authors.forename: Ryan M
authors.initials: RM
authors.lastname: O'Connell
mesh_terms: D000818:Animals; D001483:Base Sequence; D064113:CRISPR-Cas Systems; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics; D017124:Conserved Sequence; D019143:Evolution, Molecular; D023281:Genomics; D006801:Humans; D007951:Leukemia, Myeloid / Q000235:genetics / Q000473:pathology*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D008969:Molecular Sequence Data; D016159:Tumor Suppressor Protein p53 / Q000235:genetics
chemical_list: D035683:MicroRNAs; D016159:Tumor Suppressor Protein p53