title: Forward and reverse genetics through derivation of haploid mouse embryonic stem cells.
abstract: All somatic mammalian cells carry two copies of chromosomes (diploidy), whereas organisms with a single copy of their genome, such as yeast, provide a basis for recessive genetics. Here we report the generation of haploid mouse ESC lines from parthenogenetic embryos. These cells carry 20 chromosomes, express stem cell markers, and develop into all germ layers in vitro and in vivo. We also developed a reversible mutagenesis protocol that allows saturated genetic recessive screens and results in homozygous alleles. This system allowed us to generate a knockout cell line for the microRNA processing enzyme Drosha. In a forward genetic screen, we identified Gpr107 as a molecule essential for killing by ricin, a toxin being used as a bioweapon. Our results open the possibility of combining the power of a haploid genome with pluripotency of embryonic stem cells to uncover fundamental biological processes in defined cell types at a genomic scale.
authors.affiliation: IMBA, Institute of Molecular Biotechnology of the Austrian Academy of Sciences, 1030 Vienna, Austria.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Elling
authors.forename: Jasmin
authors.initials: J
authors.lastname: Taubenschmid
authors.forename: Gerald
authors.initials: G
authors.lastname: Wirnsberger
authors.forename: Ronan
authors.initials: R
authors.lastname: O'Malley
authors.forename: Simon-Pierre
authors.initials: SP
authors.lastname: Demers
authors.forename: Quentin
authors.initials: Q
authors.lastname: Vanhaelen
authors.forename: Andrey I
authors.initials: AI
authors.lastname: Shukalyuk
authors.forename: Gerald
authors.initials: G
authors.lastname: Schmauss
authors.forename: Daniel
authors.initials: D
authors.lastname: Schramek
authors.forename: Frank
authors.initials: F
authors.lastname: Schnuetgen
authors.forename: Harald
authors.initials: H
authors.lastname: von Melchner
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Ecker
authors.forename: William L
authors.initials: WL
authors.lastname: Stanford
authors.forename: Johannes
authors.initials: J
authors.lastname: Zuber
authors.forename: Alexander
authors.initials: A
authors.lastname: Stark
authors.forename: Josef M
authors.initials: JM
authors.lastname: Penninger
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D055106:Genome-Wide Association Study; D006238:Haploidy; D051379:Mice; D008810:Mice, Inbred C57BL; D010312:Parthenogenesis / Q000235:genetics; D059386:Reverse Genetics / Q000379:methods*; D012276:Ricin / Q000633:toxicity
chemical_list: D015415:Biomarkers; D012276:Ricin