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Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing
Genetically modified nonhuman primates (NHP) are useful models for biomedical research. Gene editing technologies have enabled production of target-gene knock-out (KO) NHP models. Target-gene-KO/knock-in (KI) efficiency of CRISPR/Cas9 has not been extensively investigated in marmosets. In this study...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6722079/ https://www.ncbi.nlm.nih.gov/pubmed/31481684 http://dx.doi.org/10.1038/s41598-019-49110-3 |
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author | Kumita, Wakako Sato, Kenya Suzuki, Yasuhiro Kurotaki, Yoko Harada, Takeshi Zhou, Yang Kishi, Noriyuki Sato, Kengo Aiba, Atsu Sakakibara, Yasubumi Feng, Guoping Okano, Hideyuki Sasaki, Erika |
author_facet | Kumita, Wakako Sato, Kenya Suzuki, Yasuhiro Kurotaki, Yoko Harada, Takeshi Zhou, Yang Kishi, Noriyuki Sato, Kengo Aiba, Atsu Sakakibara, Yasubumi Feng, Guoping Okano, Hideyuki Sasaki, Erika |
author_sort | Kumita, Wakako |
collection | PubMed |
description | Genetically modified nonhuman primates (NHP) are useful models for biomedical research. Gene editing technologies have enabled production of target-gene knock-out (KO) NHP models. Target-gene-KO/knock-in (KI) efficiency of CRISPR/Cas9 has not been extensively investigated in marmosets. In this study, optimum conditions for target gene modification efficacies of CRISPR/mRNA and CRISPR/nuclease in marmoset embryos were examined. CRISPR/nuclease was more effective than CRISPR/mRNA in avoiding mosaic genetic alteration. Furthermore, optimal conditions to generate KI marmoset embryos were investigated using CRISPR/Cas9 and 2 different lengths (36 nt and 100 nt) each of a sense or anti-sense single-strand oligonucleotide (ssODN). KIs were observed when CRISPR/nuclease and 36 nt sense or anti-sense ssODNs were injected into embryos. All embryos exhibited mosaic mutations with KI and KO, or imprecise KI, of c-kit. Although further improvement of KI strategies is required, these results indicated that CRISPR/Cas9 may be utilized to produce KO/KI marmosets via gene editing. |
format | Online Article Text |
id | pubmed-6722079 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-67220792019-09-17 Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing Kumita, Wakako Sato, Kenya Suzuki, Yasuhiro Kurotaki, Yoko Harada, Takeshi Zhou, Yang Kishi, Noriyuki Sato, Kengo Aiba, Atsu Sakakibara, Yasubumi Feng, Guoping Okano, Hideyuki Sasaki, Erika Sci Rep Article Genetically modified nonhuman primates (NHP) are useful models for biomedical research. Gene editing technologies have enabled production of target-gene knock-out (KO) NHP models. Target-gene-KO/knock-in (KI) efficiency of CRISPR/Cas9 has not been extensively investigated in marmosets. In this study, optimum conditions for target gene modification efficacies of CRISPR/mRNA and CRISPR/nuclease in marmoset embryos were examined. CRISPR/nuclease was more effective than CRISPR/mRNA in avoiding mosaic genetic alteration. Furthermore, optimal conditions to generate KI marmoset embryos were investigated using CRISPR/Cas9 and 2 different lengths (36 nt and 100 nt) each of a sense or anti-sense single-strand oligonucleotide (ssODN). KIs were observed when CRISPR/nuclease and 36 nt sense or anti-sense ssODNs were injected into embryos. All embryos exhibited mosaic mutations with KI and KO, or imprecise KI, of c-kit. Although further improvement of KI strategies is required, these results indicated that CRISPR/Cas9 may be utilized to produce KO/KI marmosets via gene editing. Nature Publishing Group UK 2019-09-03 /pmc/articles/PMC6722079/ /pubmed/31481684 http://dx.doi.org/10.1038/s41598-019-49110-3 Text en © The Author(s) 2019 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/. |
spellingShingle | Article Kumita, Wakako Sato, Kenya Suzuki, Yasuhiro Kurotaki, Yoko Harada, Takeshi Zhou, Yang Kishi, Noriyuki Sato, Kengo Aiba, Atsu Sakakibara, Yasubumi Feng, Guoping Okano, Hideyuki Sasaki, Erika Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing |
title | Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing |
title_full | Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing |
title_fullStr | Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing |
title_full_unstemmed | Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing |
title_short | Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing |
title_sort | efficient generation of knock-in/knock-out marmoset embryo via crispr/cas9 gene editing |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6722079/ https://www.ncbi.nlm.nih.gov/pubmed/31481684 http://dx.doi.org/10.1038/s41598-019-49110-3 |
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