ArticleNucleic acids research2012
Isolation of homozygous mutant mouse embryonic stem cells using a dual selection system.
Article in Nucleic acids research, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- CORO1C (coronin 1C) promotes autophagosome formation by coordinating branched actin network dynamics.Autophagy · 2026Article
- Aneuploid embryonic stem cells drive teratoma metastasis.Nature communications · 2024Article
- Klf4 methylated by Prmt1 restrains the commitment of primitive endoderm.Nucleic acids research · 2022Article
- Applications ofStem cells international · 2021Review
- Allicin Improves Lung Injury Induced by Sepsis via Regulation of the Toll-Like Receptor 4 (TLR4)/Myeloid Differentiation Primary Response 88 (MYD88)/Nuclear Factor kappa B (NF-κB) Pathway.Medical science monitor : international medical journal of experimental and clinical research · 2019Article
- Arrayed mutant haploid embryonic stem cell libraries facilitate phenotype-driven genetic screens.Nucleic acids research · 2017Article
- An efficient method for generation of bi-allelic null mutant mouse embryonic stem cells and its application for investigating epigenetic modifiers.Nucleic acids research · 2017Article
- Chromatin states shape insertion profiles of the piggyBac, Tol2 and Sleeping Beauty transposons and murine leukemia virus.Scientific reports · 2017Article
- Protective Effects of Methane-Rich Saline on Rats with Lipopolysaccharide-Induced Acute Lung Injury.Oxidative medicine and cellular longevity · 2017Article
- Aneuploid embryonic stem cells exhibit impaired differentiation and increased neoplastic potential.The EMBO journal · 2016Article
- CRISPR/Cas9-mediated reporter knock-in in mouse haploid embryonic stem cells.Scientific reports · 2015Article
- Application of an inducible transposon with anther culture in generation of di-haploid homologous mutants.Botanical studies · 2014Article
- A nucleolus-predominant piggyBac transposase, NP-mPB, mediates elevated transposition efficiency in mammalian cells.PloS one · 2014Article
- The piggyBac transposon displays local and distant reintegration preferences and can cause mutations at noncanonical integration sites.Molecular and cellular biology · 2013Article
- A genetic screen using the PiggyBac transposon in haploid cells identifies Parp1 as a mediator of olaparib toxicity.PloS one · 2013Article
- Interaction of Salmonella typhimurium with dendritic cells derived from pluripotent embryonic stem cells.PloS one · 2012Article
- A homozygous mutant embryonic stem cell bank applicable for phenotype-driven genetic screening.Nature methods · 2011Article
Corrections and comments
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7 authors.
Funding
Abstract
Obtaining random homozygous mutants in mammalian cells for forward genetic studies has always been problematic due to the diploid genome. With one mutation per cell, only one allele of an autosomal gene can be disrupted, and the resulting heterozygous mutant is unlikely to display a phenotype. In cells with a genetic background deficient for the Bloom's syndrome helicase, such heterozygous mutants segregate homozygous daughter cells at a low frequency due to an elevated rate of crossover following mitotic recombination between homologous chromosomes. We constructed DNA vectors that are selectable based on their copy number and used these to isolate these rare homozygous mutant cells independent of their phenotype. We use the piggyBac transposon to limit the initial mutagenesis to one copy per cell, and select for cells that have increased the transposon copy number to two or more. This yields homozygous mutants with two allelic mutations, but also cells that have duplicated the mutant chromosome and become aneuploid during culture. On average, 26% of the copy number gain events occur by the mitotic recombination pathway. We obtained homozygous cells from 40% of the heterozygous mutants tested. This method can provide homozygous mammalian loss-of-function mutants for forward genetic applications.
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