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Genome-wide barcoded transposon screen for cancer drug sensitivity in haploid mouse embryonic stem cells.

  • Author(s): Pettitt, Stephen J
  • Krastev, Dragomir B
  • Pemberton, Helen N
  • Fontebasso, Yari
  • Frankum, Jessica
  • Rehman, Farah L
  • Brough, Rachel
  • Song, Feifei
  • Bajrami, Ilirjana
  • Rafiq, Rumana
  • Wallberg, Fredrik
  • Kozarewa, Iwanka
  • Fenwick, Kerry
  • Armisen-Garrido, Javier
  • Swain, Amanda
  • Gulati, Aditi
  • Campbell, James
  • Ashworth, Alan
  • Lord, Christopher J
  • et al.

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.

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