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. 2018 May 17;8:7753. doi: 10.1038/s41598-018-25964-x

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Zygotic POU5F1 is required for expanded blastocyst formation in in vitro fertilized bovine embryos. (A) Embryo development following peri-zygotic injection of CRISPR/Cas9 targeting the POU5F1 locus was determined at the 2C (cell), 8–16C and blastocyst stages (D7.5) of oocytes fertilized with frozen-thawed bull sperm. (B) Immunofluorescent detection of POU5F1 expression was determined in D7.5 morulae from the targeted knockout (TKO) and injected control (IC) groups. (C) Developmentally advanced embryos from the TKO and IC groups were evaluated for POU5F1 (green) and CDX2 (red) expression. Arrows indicate a POU5F1 negative morula. ^a,bDifferent letters indicate significant differences (P < 0.05). Scale bar = 100 µm.