Unknown

Dataset Information

0

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.

SUBMITTER: Pettitt SJ 

PROVIDER: S-EPMC5332012 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

Json Xml
altmetric image

Publications

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 the  ...[more]

Similar Datasets

Unknown Germline potential of parthenogenetic haploid mouse embryonic stem cells.
| S-EPMC3424041 | biostudies-literature
Unknown Derivation and maintenance of mouse haploid embryonic stem cells.
| S-EPMC6997032 | biostudies-literature
Unknown Derivation of haploid embryonic stem cells from mouse embryos.
| S-EPMC3209452 | biostudies-literature
Unknown Genome-wide genetic screening with chemically mutagenized haploid embryonic stem cells.
| S-EPMC5164930 | biostudies-literature
Unknown A Genome-wide Haploid Genetic Screen Identifies Regulators of Glutathione Abundance and Ferroptosis Sensitivity.
| S-EPMC6424331 | biostudies-literature
Unknown A Genome-Wide CRISPR Screen Identifies Factors Regulating Pluripotency Exit in Mouse Embryonic Stem Cells.
| S-EPMC9331787 | biostudies-literature
Unknown Forward and reverse genetics through derivation of haploid mouse embryonic stem cells.
| S-EPMC4008724 | biostudies-literature
Unknown CRISPR/Cas9-mediated reporter knock-in in mouse haploid embryonic stem cells.
| S-EPMC4454075 | biostudies-literature
Unknown Transcriptome profiles and chromatin states in mouse androgenetic haploid embryonic stem cells.
| S-EPMC10472531 | biostudies-literature
Unknown A reversible haploid mouse embryonic stem cell biobank resource for functional genomics.
| S-EPMC6235111 | biostudies-literature