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

Item Type:	Article
Title:	A reversible haploid mouse embryonic stem cell biobank resource for functional genomics
Creators Name:	Elling, U. and Wimmer, R.A. and Leibbrandt, A. and Burkard, T. and Michlits, G. and Leopoldi, A. and Micheler, T. and Abdeen, D. and Zhuk, S. and Aspalter, I.M. and Handl, C. and Liebergesell, J. and Hubmann, M. and Husa, A.M. and Kinzer, M. and Schuller, N. and Wetzel, E. and van de Loo, N. and Martinez, J.A.Z. and Estoppey, D. and Riedl, R. and Yang, F. and Fu, B. and Dechat, T. and Ivics, Z. and Agu, C.A. and Bell, O. and Blaas, D. and Gerhardt, H. and Hoepfner, D. and Stark, A. and Penninger, J.M.
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.
Keywords:	Biological Specimen Banks, Blood Vessels, Cell Lineage, Common Cold, Genes, Essential, Genetic Testing, Genomics, HEK293 Cells, Haploidy, Homozygote, Mouse Embryonic Stem Cells, Mutation, Neovascularization, Physiologic, Phospholipases A2, Calcium-Independent, Rhinovirus, Animals, Mice
Source:	Nature
ISSN:	0028-0836
Publisher:	Nature Publishing Group
Volume:	550
Number:	7674
Page Range:	114-118
Date:	5 October 2017
Official Publication:	https://doi.org/10.1038/nature24027
External Fulltext:	View full text on PubMed Central
PubMed:	View item in PubMed

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