title: Discovery of cancer drug targets by CRISPR-Cas9 screening of protein domains.
abstract: CRISPR-Cas9 genome editing technology holds great promise for discovering therapeutic targets in cancer and other diseases. Current screening strategies target CRISPR-Cas9-induced mutations to the 5' exons of candidate genes, but this approach often produces in-frame variants that retain functionality, which can obscure even strong genetic dependencies. Here we overcome this limitation by targeting CRISPR-Cas9 mutagenesis to exons encoding functional protein domains. This generates a higher proportion of null mutations and substantially increases the potency of negative selection. We also show that the magnitude of negative selection can be used to infer the functional importance of individual protein domains of interest. A screen of 192 chromatin regulatory domains in murine acute myeloid leukemia cells identifies six known drug targets and 19 additional dependencies. A broader application of this approach may allow comprehensive identification of protein domains that sustain cancer cells and are suitable for drug targeting.
authors.affiliation: 1] Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA. [2] Molecular and Cellular Biology Program, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Junwei
authors.initials: J
authors.lastname: Shi
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Wang
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Milazzo
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.
authors.forename: Zihua
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.
authors.forename: Justin B
authors.initials: JB
authors.lastname: Kinney
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Vakoc
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D045744:Cell Line, Tumor; D055808:Drug Discovery; D005091:Exons / Q000235:genetics; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy / Q000235:genetics*; D051379:Mice; D058990:Molecular Targeted Therapy; D016296:Mutagenesis / Q000235:genetics; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics*; D017434:Protein Structure, Tertiary / Q000235:genetics; D017393:RNA Editing / Q000235:genetics
chemical_list: D009363:Neoplasm Proteins