ArticleBMC cell biology2016
Aneuploidy screening of embryonic stem cell clones by metaphase karyotyping and droplet digital polymerase chain reaction.
Article in BMC cell biology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 40 citations in OpenAlex.
- Chromosome X dosage modulates development of aneuploidy in genetically diverse mouse embryonic stem cells.Stem cell reports · 2026Article
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- Development of HPV16 mouse and dog models for more accurate prediction of human vaccine efficacy.Laboratory animal research · 2023Article
- Excess of guide RNA reduces knockin efficiency and drastically increases on-target large deletions.iScience · 2023Article
- Cytosine base editors induce off-target mutations and adverse phenotypic effects in transgenic mice.Nature communications · 2023Article
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- Increased On-Target Rate and Risk of Concatemerization after CRISPR-Enhanced Targeting in ES Cells.Genes · 2023Article
- Conventional and Spectral Karyotyping of Murine Cerebellar Granule Neuron Progenitors.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Ts66Yah, a mouse model of Down syndrome with improved construct and face validity.Disease models & mechanisms · 2022Article
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- Dynamic Runx1 chromatin boundaries affect gene expression in hematopoietic development.Nature communications · 2022Article
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- High-throughput genotyping of high-homology mutant mouse strains by next-generation sequencing.Methods (San Diego, Calif.) · 2021Article
- Universal Southern blot protocol with cold or radioactive probes for the validation of alleles obtained by homologous recombination.Methods (San Diego, Calif.) · 2021Article
- All models are wrong, but some are useful: Establishing standards for stem cell-based embryo models.Stem cell reports · 2021Review
- Article
- DNA barcoding reveals that injected transgenes are predominantly processed by homologous recombination in mouse zygote.Nucleic acids research · 2020Article
- Application of long single-stranded DNA donors in genome editing: generation and validation of mouse mutants.BMC biology · 2018Article
- Next-generation mammalian genetics toward organism-level systems biology.NPJ systems biology and applications · 2017Review
Corrections and comments
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Authors and funding
15 authors at 2 institutions in 2 countries.
Funding
Abstract
backgroundKaryotypic integrity is essential for the successful germline transmission of alleles mutated in embryonic stem (ES) cells. Classical methods for the identification of aneuploidy involve cytological analyses that are both time consuming and require rare expertise to identify mouse chromosomes.
resultsAs part of the International Mouse Phenotyping Consortium, we gathered data from over 1,500 ES cell clones and found that the germline transmission (GLT) efficiency of clones is compromised when over 50 % of cells harbour chromosome number abnormalities. In JM8 cells, chromosomes 1, 8, 11 or Y displayed copy number variation most frequently, whilst the remainder generally remain unchanged. We developed protocols employing droplet digital polymerase chain reaction (ddPCR) to accurately quantify the copy number of these four chromosomes, allowing efficient triage of ES clones prior to microinjection. We verified that assessments of aneuploidy, and thus decisions regarding the suitability of clones for microinjection, were concordant between classical cytological and ddPCR-based methods. Finally, we improved the method to include assay multiplexing so that two unstable chromosomes are counted simultaneously (and independently) in one reaction, to enhance throughput and further reduce the cost.
conclusionWe validated a PCR-based method as an alternative to classical karyotype analysis. This technique enables laboratories that are non-specialist, or work with large numbers of clones, to precisely screen ES cells for the most common aneuploidies prior to microinjection to ensure the highest level of germline transmission potential. The application of this method allows early exclusion of aneuploid ES cell clones in the ES cell to mouse conversion process, thus improving the chances of obtaining germline transmission and reducing the number of animals used in failed microinjection attempts. This method can be applied to any other experiments that require accurate analysis of the genome for copy number variation (CNV).
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.