A reversible haploid mouse embryonic stem cell biobank resource for functional genomics

被引：56

作者：

Elling, Ulrich ^{[1
]}

Wimmer, Reiner A. ^{[1
]}

Leibbrandt, Andreas ^{[1
]}

Urkard, Thomas B. ^{[1
]}

Michlits, Georg ^{[1
]}

Leopoldi, Alexandra ^{[1
]}

Micheler, Thomas ^{[2
]}

Abdeen, Dana ^{[1
]}

Zhuk, Sergei ^{[1
]}

Aspalter, Irene M. ^{[3
,4
]}

Handl, Cornelia ^{[1
]}

Liebergesell, Julia ^{[1
]}

Hubmann, Maria ^{[1
]}

Husa, Anna-Maria ^{[1
]}

Kinzer, Manuela ^{[1
]}

Schuller, Nicole ^{[1
]}

Wetzel, Ellen ^{[1
]}

van de Loo, Nina ^{[1
]}

Zepeda Martinez, Jorge Arturo ^{[1
]}

Estoppey, David ^{[5
]}

Riedl, Ralph ^{[5
]}

Yang, Fengtang ^{[6
]}

Fu, Beiyuan ^{[6
]}

Dechat, Thomas ^{[7
]}

Ivics, Zoltan ^{[8
]}

Agu, Chukwuma A. ^{[1
]}

Bell, Oliver ^{[1
]}

Blaas, Dieter ^{[7
]}

Gerhardt, Holger ^{[9
,10
,11
]}

Hoepfner, Dominic ^{[5
]}

Stark, Alexander ^{[12
]}

Penninger, Josef M. ^{[1
]}

机构：

[1] Austrian Acad Sci IMBA, Inst Mol Biotechnol, Vienna Bioctr VBC, Dr Bohr Gasse 3, Vienna, Austria

[2] Vienna Bioctr VBC, Vienna Bioctr Core Facil, Dr Bohr Gasse 3, Vienna, Austria

[3] UCL, MRC Lab Mol Cell Biol, London, England

[4] UCL, Inst Phys Living Syst, London, England

[5] Novartis Inst BioMed Res, Basel, Switzerland

[6] Wellcome Trust Sanger Inst, Cambridge CB10 1SA, England

[7] Med Univ Vienna, Max F Perutz Labs, Dr Bohr Gasse 9, Vienna, Austria

[8] Paul Ehrlich Inst, Paul Ehrlich Str 51-59, D-63225 Langen, Germany

[9] Helmholtz Assoc, Max Delbruck Ctr Mol Med, Berlin, Germany

[10] German Ctr Cardiovasc Res, Berlin, Germany

[11] Berlin Inst Hlth, Berlin, Germany

[12] Vienna Bioctr VBC, Res Inst Mol Pathol IMP, Dr Bohr Gasse 7, A-1030 Vienna, Austria

来源：

NATURE | 2017年 / 550卷 / 7674期

基金：

奥地利科学基金会;

关键词：

DENSITY-LIPOPROTEIN RECEPTOR; PREIMPLANTATION DEVELOPMENT; MAMMALIAN-CELLS; ESSENTIAL GENES; RNAI SCREENS; ANGIOGENESIS; IDENTIFICATION; HETEROGENEITY; MUTAGENESIS; TRANSPOSON;

D O I：

10.1038/nature24027

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

The ability to directly uncover the contributions of genes to a given phenotype is fundamental for biology research. However, ostensibly homogeneous cell populations exhibit large clonal variance(1,2) that can confound analyses and undermine reproducibility(3). Here we used genome-saturated mutagenesis to create a biobank of over 100,000 individual haploid mouse embryonic stem (mES) cell lines targeting 16,970 genes with genetically barcoded, conditional and reversible mutations. This Haplobank is, to our knowledge, the largest resource of hemi/homozygous mutant mES cells to date and is available to all researchers. Reversible mutagenesis overcomes clonal variance by permitting functional annotation of the genome directly in sister cells. We use the Haplobank in reverse genetic screens to investigate the temporal resolution of essential genes in mES cells, and to identify novel genes that control sprouting angiogenesis and lineage specification of blood vessels. Furthermore, a genome-wide forward screen with Haplobank identified PLA2G16 as a host factor that is required for cytotoxicity by rhinoviruses, which cause the common cold. Therefore, clones from the Haplobank combined with the use of reversible technologies enable high-throughput, reproducible, functional annotation of the genome.

引用

页码：114 / +

页数：27

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