Highly Efficient CRISPR-Associated Protein 9 Ribonucleoprotein-Based Genome Editing in Euglena gracilis
Summary: Euglena gracilis, a unicellular phytoflagellate microalga, is a promising biomaterial for foods, feeds, and biofuels. However, targeted mutagenesis in this species has been a long-standing challenge. We recently developed a transgene-free, highly efficient, genome editing method for E. grac...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-06-01
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| Series: | STAR Protocols |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166720300101 |
| _version_ | 1828955522510880768 |
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| author | Toshihisa Nomura Mizuki Yoshikawa Kengo Suzuki Keiichi Mochida |
| author_facet | Toshihisa Nomura Mizuki Yoshikawa Kengo Suzuki Keiichi Mochida |
| author_sort | Toshihisa Nomura |
| collection | DOAJ |
| description | Summary: Euglena gracilis, a unicellular phytoflagellate microalga, is a promising biomaterial for foods, feeds, and biofuels. However, targeted mutagenesis in this species has been a long-standing challenge. We recently developed a transgene-free, highly efficient, genome editing method for E. gracilis using CRISPR/Cas9 ribonucleoproteins (RNPs). Our method achieved mutagenesis rates of approximately 80% or more through an electroporation-based direct delivery of Cas9 RNPs. Therefore, this method is suitable for basic research and industrial applications, such as the breeding of Euglena.For complete details on the use and execution of this protocol, please refer to Nomura et al. (2019). |
| first_indexed | 2024-12-14T07:51:44Z |
| format | Article |
| id | doaj.art-36f0742036a240c7b1dc6c46a6d9af42 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-14T07:51:44Z |
| publishDate | 2020-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-36f0742036a240c7b1dc6c46a6d9af422022-12-21T23:10:40ZengElsevierSTAR Protocols2666-16672020-06-0111100023Highly Efficient CRISPR-Associated Protein 9 Ribonucleoprotein-Based Genome Editing in Euglena gracilisToshihisa Nomura0Mizuki Yoshikawa1Kengo Suzuki2Keiichi Mochida3RIKEN Center for Sustainable Resource Science, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan; RIKEN Baton Zone Program, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan; Corresponding authorRIKEN Baton Zone Program, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, JapanRIKEN Baton Zone Program, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan; euglena Co., Ltd., 5-33-1 Shiba, Minato-ku, Tokyo 108-0014, JapanRIKEN Center for Sustainable Resource Science, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan; RIKEN Baton Zone Program, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan; Kihara Institute for Biological Research, Yokohama City University, 641-12 Maioka-cho, Totsuka-ku, Yokohama, Kanagawa 244-0813, Japan; Graduate School of Nanobioscience, Yokohama City University, 1-7-29 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan; Institute of Plant Science and Resources, Okayama University, 2-20-1 Chuo, Kurashiki, Okayama 710-0046, Japan; Corresponding authorSummary: Euglena gracilis, a unicellular phytoflagellate microalga, is a promising biomaterial for foods, feeds, and biofuels. However, targeted mutagenesis in this species has been a long-standing challenge. We recently developed a transgene-free, highly efficient, genome editing method for E. gracilis using CRISPR/Cas9 ribonucleoproteins (RNPs). Our method achieved mutagenesis rates of approximately 80% or more through an electroporation-based direct delivery of Cas9 RNPs. Therefore, this method is suitable for basic research and industrial applications, such as the breeding of Euglena.For complete details on the use and execution of this protocol, please refer to Nomura et al. (2019).http://www.sciencedirect.com/science/article/pii/S2666166720300101 |
| spellingShingle | Toshihisa Nomura Mizuki Yoshikawa Kengo Suzuki Keiichi Mochida Highly Efficient CRISPR-Associated Protein 9 Ribonucleoprotein-Based Genome Editing in Euglena gracilis STAR Protocols |
| title | Highly Efficient CRISPR-Associated Protein 9 Ribonucleoprotein-Based Genome Editing in Euglena gracilis |
| title_full | Highly Efficient CRISPR-Associated Protein 9 Ribonucleoprotein-Based Genome Editing in Euglena gracilis |
| title_fullStr | Highly Efficient CRISPR-Associated Protein 9 Ribonucleoprotein-Based Genome Editing in Euglena gracilis |
| title_full_unstemmed | Highly Efficient CRISPR-Associated Protein 9 Ribonucleoprotein-Based Genome Editing in Euglena gracilis |
| title_short | Highly Efficient CRISPR-Associated Protein 9 Ribonucleoprotein-Based Genome Editing in Euglena gracilis |
| title_sort | highly efficient crispr associated protein 9 ribonucleoprotein based genome editing in euglena gracilis |
| url | http://www.sciencedirect.com/science/article/pii/S2666166720300101 |
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