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Nature. 2017 Oct 5;550(7674):114-118. doi: 10.1038/nature24027. Epub 2017 Sep 27.

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

Author information

1
Institute of Molecular Biotechnology of the Austrian Academy of Science (IMBA), Vienna Biocenter (VBC), Dr. Bohr Gasse 3, Vienna, Austria.
2
Vienna Biocenter Core Facilities, Vienna Biocenter (VBC), Dr. Bohr Gasse 3, Vienna, Austria.
3
MRC Laboratory for Molecular Cell Biology and Institute for the Physics of Living Systems, University College London, London, UK.
4
Novartis Institutes for BioMedical Research, Basel, Switzerland.
5
Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK.
6
Max F. Perutz Laboratories, Medical University of Vienna, Dr. Bohr Gasse 9, Vienna, Austria.
7
Paul Ehrlich Institut, Paul Ehrlich Strasse 51-59, 63225 Langen, Germany.
8
Max-Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany.
9
German Center for Cardiovascular Research, Berlin, Germany.
10
Berlin Institute of Health, Berlin, Germany.
11
Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), Dr. Bohr Gasse 7, 1030 Vienna, Austria.

Abstract

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 that can confound analyses and undermine reproducibility. 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.

PMID:
28953874
PMCID:
PMC6235111
DOI:
10.1038/nature24027
[Indexed for MEDLINE]
Free PMC Article

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