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.
journal_name
Sci Datajournal_title
Scientific dataauthors
Pettitt SJ,Krastev DB,Pemberton HN,Fontebasso Y,Frankum J,Rehman FL,Brough R,Song F,Bajrami I,Rafiq R,Wallberg F,Kozarewa I,Fenwick K,Armisen-Garrido J,Swain A,Gulati A,Campbell J,Ashworth A,Lord CJdoi
10.1038/sdata.2017.20subject
Has Abstractpub_date
2017-03-01 00:00:00pages
170020issn
2052-4463pii
sdata201720journal_volume
4pub_type
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