ArticleNucleic acids research2017
Arrayed mutant haploid embryonic stem cell libraries facilitate phenotype-driven genetic screens.
Article in Nucleic acids research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 43 citations in OpenAlex.
- CORO1C (coronin 1C) promotes autophagosome formation by coordinating branched actin network dynamics.Autophagy · 2026Article
- Haploid-genetic screening of trophectoderm specification identifiesScience advances · 2023Article
- BCL2 is a major regulator of haploidy maintenance in murine embryonic stem cells.Cell proliferation · 2023Article
- Genome-scale screening in a rat haploid system identifies Thop1 as a modulator of pluripotency exit.Cell proliferation · 2022Article
- hnRNPLL controls pluripotency exit of embryonic stem cells by modulating alternative splicing of Tbx3 and Bptf.The EMBO journal · 2021Article
- A Genetic Screen Identifies Etl4-Deficiency Capable of Stabilizing the Haploidy in Embryonic Stem Cells.Stem cell reports · 2021Article
- Applications ofStem cells international · 2021Review
- Advances in high-throughput methods for the identification of virus receptors.Medical microbiology and immunology · 2020Review
- Current advances in haploid stem cells.Protein & cell · 2020Review
- The milestone of genetic screening: Mammalian haploid cells.Computational and structural biotechnology journal · 2020Review
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
Abstract
Forward genetic screens using mammalian embryonic stem (ES) cells have identified genes required for numerous cellular processes. However, loss-of-function screens are more difficult to conduct in diploid cells because, in most cases, both alleles of a gene must be mutated to exhibit a phenotype. Recently, mammalian haploid ES cell lines were successfully established and applied to several recessive genetic screens. However, all these screens were performed in mixed pools of mutant cells and were mainly based on positive selection. In general, negative screening is not easy to apply to these mixed pools, although quantitative deep sequencing of mutagen insertions can help to identify some 'missing' mutants. Moreover, the interplay between different mutant cells in the mixed pools would interfere with the readout of the screens. Here, we developed a method for rapidly generating arrayed haploid mutant libraries in which the proportion of homozygous mutant clones can reach 85%. After screening thousands of individual mutant clones, we identified a number of novel factors required for the onset of differentiation in ES cells. A negative screen was also conducted to discover mutations conferring cells with increased sensitivity to DNA double-strand breaks induced by the drug doxorubicin. Both of these screens illustrate the value of this system.
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Registered trials
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