Genome editing reveals a role for OCT4 in human embryogenesis

被引：0
作者：
Norah M. E. Fogarty
Afshan McCarthy
Kirsten E. Snijders
Benjamin E. Powell
Nada Kubikova
Paul Blakeley
Rebecca Lea
Kay Elder
Sissy E. Wamaitha
Daesik Kim
Valdone Maciulyte
Jens Kleinjung
Jin-Soo Kim
Dagan Wells
Ludovic Vallier
Alessandro Bertero
James M. A. Turner
Kathy K. Niakan
机构：
[1] Human Embryo and Stem Cell Laboratory,NIHR Cambridge Biomedical Research Centre hIPSC Core Facility Department of Surgery
[2] The Francis Crick Institute,Nuffield Department of Obstetrics and Gynaecology
[3] NIHR Cambridge Biomedical Research Centre hIPSC Core Facility,Department of Chemistry
[4] University of Cambridge,Department of Surgery University of Cambridge
[5] Cambridge Biomedical Campus,Department of Pathology
[6] Sex Chromosome Biology Laboratory,undefined
[7] The Francis Crick Institute,undefined
[8] University of Oxford,undefined
[9] John Radcliffe Hospital,undefined
[10] Bourn Hall Clinic,undefined
[11] Seoul National University,undefined
[12] Bioinformatics Facility,undefined
[13] The Francis Crick Institute,undefined
[14] Center for Genome Engineering,undefined
[15] Institute for Basic Science,undefined
[16] Wellcome Trust Sanger Institute,undefined
[17] Wellcome Genome Campus,undefined
[18] Wellcome Trust and MRC Cambridge Stem Cell Institute and Biomedical Research Centre,undefined
[19] Anne McLaren Laboratory,undefined
[20] University of Cambridge,undefined
[21] University of Washington,undefined
来源：
Nature | 2017年 / 550卷
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摘要：
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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页码：67 / 73
页数：6
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