Genome-wide genetic screening with chemically mutagenized haploid embryonic stem cells.
In model organisms, classical genetic screening via random mutagenesis provides key insights into the molecular bases of genetic interactions, helping to define synthetic lethality, synthetic viability and drug-resistance mechanisms. The limited genetic tractability of diploid mammalian cells, however, precludes this approach. Here, we demonstrate the feasibility of classical genetic screening in mammalian systems by using haploid cells, chemical mutagenesis and next-generation sequencing, providing a new tool to explore mammalian genetic interactions.
The Wellcome Trust and Cancer Research UK Gurdon Institute, and Department of Biochemistry, University of Cambridge, Cambridge, UK.|The Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK.
Josep V
http://orcid.org/0000-0002-7797-2583
JV
Forment
The Wellcome Trust and Cancer Research UK Gurdon Institute, and Department of Biochemistry, University of Cambridge, Cambridge, UK.|The Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK.
Mareike
http://orcid.org/0000-0001-9747-2327
M
Herzog
The Wellcome Trust and Cancer Research UK Gurdon Institute, and Department of Biochemistry, University of Cambridge, Cambridge, UK.
Julia
J
Coates
Ludwig Institute for Cancer Research Ltd. and Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Oxford, UK.
Tomasz
T
Konopka
Ludwig Institute for Cancer Research Ltd. and Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Oxford, UK.
Bianca V
BV
Gapp
Ludwig Institute for Cancer Research Ltd. and Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Oxford, UK.|Research Center for Molecular Medicine of the Austrian Academy of Sciences (CeMM), Vienna, Austria.
Sebastian M
SM
Nijman
The Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK.
David J
DJ
Adams
The Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK.
Thomas M
TM
Keane
The Wellcome Trust and Cancer Research UK Gurdon Institute, and Department of Biochemistry, University of Cambridge, Cambridge, UK.|The Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK.
Stephen P
http://orcid.org/0000-0001-9317-7937
SP
Jackson
D000818:Animals; D002460:Cell Line; D005820:Genetic Testing; D016678:Genome / Q000187:drug effects* / Q000235:genetics*; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000187:drug effects* / Q000378:metabolism*; D016296:Mutagenesis / Q000187:drug effects*