title: Genome-wide barcoded transposon screen for cancer drug sensitivity in haploid mouse embryonic stem cells.
abstract: We describe a screen for cellular response to drugs that makes use of haploid embryonic stem cells. We generated ten libraries of mutants with piggyBac gene trap transposon integrations, totalling approximately 100,000 mutant clones. Random barcode sequences were inserted into the transposon vector to allow the number of cells bearing each insertion to be measured by amplifying and sequencing the barcodes. These barcodes were associated with their integration sites by inverse PCR. We exposed these libraries to commonly used cancer drugs and profiled changes in barcode abundance by Ion Torrent sequencing in order to identify mutations that conferred sensitivity. Drugs tested included conventional chemotherapeutics as well as targeted inhibitors of topoisomerases, poly(ADP-ribose) polymerase (PARP), Hsp90 and WEE1.
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Pettitt
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Dragomir B
authors.initials: DB
authors.lastname: Krastev
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Helen N
authors.initials: HN
authors.lastname: Pemberton
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Yari
authors.initials: Y
authors.lastname: Fontebasso
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Jessica
authors.initials: J
authors.lastname: Frankum
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Farah L
authors.initials: FL
authors.lastname: Rehman
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Rachel
authors.initials: R
authors.lastname: Brough
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Feifei
authors.initials: F
authors.lastname: Song
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Ilirjana
authors.initials: I
authors.lastname: Bajrami
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Rumana
authors.initials: R
authors.lastname: Rafiq
authors.affiliation: Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Fredrik
authors.initials: F
authors.lastname: Wallberg
authors.affiliation: Tumour Profiling Unit, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Iwanka
authors.initials: I
authors.lastname: Kozarewa
authors.affiliation: Tumour Profiling Unit, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Kerry
authors.initials: K
authors.lastname: Fenwick
authors.affiliation: Tumour Profiling Unit, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Javier
authors.initials: J
authors.lastname: Armisen-Garrido
authors.affiliation: Tumour Profiling Unit, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Amanda
authors.initials: A
authors.lastname: Swain
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Aditi
authors.initials: A
authors.lastname: Gulati
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: James
authors.initials: J
authors.lastname: Campbell
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Alan
authors.initials: A
authors.lastname: Ashworth
authors.affiliation: The CRUK Gene Function Laboratory, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.|Breast Cancer Now Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Lord
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D004251:DNA Transposable Elements; D055106:Genome-Wide Association Study; D006238:Haploidy; D051379:Mice; D000066450:Mouse Embryonic Stem Cells; D009369:Neoplasms / Q000188:drug therapy / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D004251:DNA Transposable Elements