title: Live imaging RNAi screen reveals genes essential for meiosis in mammalian oocytes.
abstract: During fertilization, an egg and a sperm fuse to form a new embryo. Eggs develop from oocytes in a process called meiosis. Meiosis in human oocytes is highly error-prone, and defective eggs are the leading cause of pregnancy loss and several genetic disorders such as Down's syndrome. Which genes safeguard accurate progression through meiosis is largely unclear. Here we develop high-content phenotypic screening methods for the systematic identification of mammalian meiotic genes. We targeted 774 genes by RNA interference within follicle-enclosed mouse oocytes to block protein expression from an early stage of oocyte development onwards. We then analysed the function of several genes simultaneously by high-resolution imaging of chromosomes and microtubules in live oocytes and scored each oocyte quantitatively for 50 phenotypes, generating a comprehensive resource of meiotic gene function. The screen generated an unprecedented annotated data set of meiotic progression in 2,241 mammalian oocytes, which allowed us to analyse systematically which defects are linked to abnormal chromosome segregation during meiosis, identifying progression into anaphase with misaligned chromosomes as well as defects in spindle organization as risk factors. This study demonstrates how high-content screens can be performed in oocytes, and allows systematic studies of meiosis in mammals.
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH, United Kingdom.
authors.forename: Sybille
authors.initials: S
authors.lastname: Pfender
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH, United Kingdom.
authors.forename: Vitaliy
authors.initials: V
authors.lastname: Kuznetsov
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH, United Kingdom.
authors.forename: Michał
authors.initials: M
authors.lastname: Pasternak
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH, United Kingdom.
authors.forename: Thomas
authors.initials: T
authors.lastname: Tischer
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH, United Kingdom.
authors.forename: Balaji
authors.initials: B
authors.lastname: Santhanam
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH, United Kingdom.
authors.forename: Melina
authors.initials: M
authors.lastname: Schuh
mesh_terms: D000705:Anaphase / Q000235:genetics; D000782:Aneuploidy; D000818:Animals; D020090:Chromosome Segregation / Q000235:genetics; D002875:Chromosomes / Q000235:genetics / Q000378:metabolism; D054637:Dual-Specificity Phosphatases / Q000235:genetics; D005260:Female; D042822:Genomic Instability / Q000235:genetics; D008297:Male; D008540:Meiosis / Q000235:genetics*; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000235:genetics; D008870:Microtubules / Q000235:genetics / Q000378:metabolism; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism*; D006080:Ovarian Follicle / Q000166:cytology; D010641:Phenotype; D014176:Protein Biosynthesis; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D034622:RNA Interference; D008941:Spindle Apparatus / Q000235:genetics / Q000378:metabolism
chemical_list: D008869:Microtubule-Associated Proteins; D017346:Protein Serine-Threonine Kinases; C585164:greatwall protein, mouse; D054637:Dual-Specificity Phosphatases; C529383:Dusp7 protein, mouse