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Genome editing reveals a role for OCT4 in human embryogenesis
Open accessAbstract
Despite their fundamental biological and clinical importance, the molecular mechanisms that regulate the first cell fate decisions in the human embryo are not well understood. Here we use CRISPR-Cas9-mediated genome editing to investigate the function of the pluripotency transcription factor OCT4 during human embryogenesis. We identified an efficient OCT4-targeting guide RNA using an inducible human embryonic stem cell-based system and microinjection of mouse zygotes. Using these refined methods, we efficiently and specifically targeted the gene encoding OCT4 (POU5F1) in diploid human zygotes and found that blastocyst development was compromised. Transcriptomics analysis revealed that, in POU5F1-null cells, gene expression was downregulated not only for extra-embryonic trophectoderm genes, such as CDX2, but also for regulators of the pluripotent epiblast, including NANOG. By contrast, Pou5f1-null mouse embryos maintained the expression of orthologous genes, and blastocyst development was established, but maintenance was compromised. We conclude that CRISPR-Cas9-mediated genome editing is a powerful method for investigating gene function in the context of human development.
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“Germline editing has been used for years in animals, including mice, rats, monkeys and many others, and experiments show that it can be done in both nonviable or viable human embryos as well 110-113 .”
From The promise and challenge of therapeutic genome editing (2020) · cites this paper as [110]
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“A report of inaccurate CRISPR-Cas9based genome editing in non-viable human embryos 110 was not substantiated by later publications, but the mechanism by which double-stranded DNA breaks are repaired in early human embryos remains under debate.”
From The promise and challenge of therapeutic genome editing (2020) · cites this paper as [110]
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“Moreover, we identified that 57 of the 137 successfully genotyped samples (42%) exhibited a homozygous wild-type genotype based on PCR amplification of a short genomic fragment (17).”
From Frequent loss of heterozygosity in CRISPR-Cas9–edited early human embryos (2021) · cites this paper as [17]
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“We generated a number of single-cell amplified genomic DNA (gDNA) samples for genotyping and confirmed on-target genome editing in all microinjected embryos and a stereotypic insertion/deletion (indel) pattern of mutations with the majority of samples exhibiting a 2bp deletion (17).”
From Frequent loss of heterozygosity in CRISPR-Cas9–edited early human embryos (2021) · cites this paper as [17]
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“This corroborates our previous findings in human embryos targeted post-fertilisation, where we noted a stereotypic pattern to the type of indel mutations and speculated that this was likely due MMEJ (17).”
From Frequent loss of heterozygosity in CRISPR-Cas9–edited early human embryos (2021) · cites this paper as [17]
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Metadata
- Authors
- Fogarty, N. M. E., McCarthy, A., Snijders, K. E., Powell, B. E., Kubikova, N., Blakeley, P., Lea, R., Elder, K., Wamaitha, S. E., Kim, D., Maciulyte, V., Kleinjung, J., Kim, J.-S., Wells, D., Vallier, L., Bertero, A., Turner, J. M. A., & Niakan, K. K.
- Affiliations
- The Francis Crick Institute (GB)
- University of Cambridge (GB)
- John Radcliffe Hospital (GB)
- Bourn Hall Clinic (GB)
- Seoul National University (KR)
- Wellcome/MRC Cambridge Stem Cell Institute (GB)
- Corresponding author
- Niakan, Kathy K.
- ORCID
- Venue
- Nature
- Published
- 2017-09-20
- Volume
- 550
- Issue
- 7674
- Pages
- 67-73
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC5815497
- OpenAlex
- W2759601393
- Cited by
- 392
- References
- 51
- Citations by year
- 2026: 16 · 2025: 24 · 2024: 34 · 2023: 45 · 2022: 35 · 2021: 50 · 2020: 50 · 2019: 62 · 2018: 62 · 2017: 13
- FWCI
- 20.88
- Type
- journal article
- Language
- en
- Update notices
- erratum (2017-10-04)
- Open access
- bronze
- doi.org
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- Research Portal (King's College London) (publishedVersion, other-oa)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Institutional Research Information System University of Turin (University of Turin) (submittedVersion, other-oa)
- Apollo (University of Cambridge) (submittedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- Figshare (submittedVersion, cc-by)
- Seoul National University Open Repository (Seoul National University) (submittedVersion)
- Apollo
- Funders
- Wellcome Trust (FC001193, FC001120)
- Francis Crick Institute (10120, 10002, FC001193, 10193, FC001120)
- Cancer Research UK (FC001120, FC001193)
- National Institute for Health and Care Research
- British Heart Foundation (FS/11/77/39327)
- University of Oxford
- Institute for Basic Science (IBSR021-D1, R021-D1)
- Medical Research Council (PSAG028, FC001193, MC_PC_12009, FC001120)
- Clarendon Fund
- NIHR Cambridge Biomedical Research Centre
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Keywords
- Oct4, human embryonic development, blastocyst development, pluripotency transcription factors, trophectoderm, Nanog, epiblast, CRISPR/Cas9 genome editing, human zygotes, guide RNA design, placental development
- MeSH terms
- Human Embryonic Stem Cells, Nanog Homeobox Protein, Gene Editing, Animals, Blastocyst, Ectoderm, Embryo, Mammalian, Female, Germ Layers, Humans, Male, Substrate Specificity, Zygote, Gene Expression Regulation, Developmental, Cell Lineage, Embryonic Development, Octamer Transcription Factor-3, Mice, CRISPR-Cas Systems
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