Highly Efficient CRISPR-Associated Protein 9 Ribonucleoprotein-Based Genome Editing in Euglena gracilis

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...

Full description

Bibliographic Details
Main Authors: Toshihisa Nomura, Mizuki Yoshikawa, Kengo Suzuki, Keiichi Mochida
Format: Article
Language:English
Published: Elsevier 2020-06-01
Series:STAR Protocols
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166720300101
id doaj-36f0742036a240c7b1dc6c46a6d9af42
record_format Article
spelling doaj-36f0742036a240c7b1dc6c46a6d9af422020-11-25T04:01:29ZengElsevierSTAR 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
collection DOAJ
language English
format Article
sources DOAJ
author Toshihisa Nomura
Mizuki Yoshikawa
Kengo Suzuki
Keiichi Mochida
spellingShingle Toshihisa Nomura
Mizuki Yoshikawa
Kengo Suzuki
Keiichi Mochida
Highly Efficient CRISPR-Associated Protein 9 Ribonucleoprotein-Based Genome Editing in Euglena gracilis
STAR Protocols
author_facet Toshihisa Nomura
Mizuki Yoshikawa
Kengo Suzuki
Keiichi Mochida
author_sort Toshihisa Nomura
title 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_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_sort highly efficient crispr-associated protein 9 ribonucleoprotein-based genome editing in euglena gracilis
publisher Elsevier
series STAR Protocols
issn 2666-1667
publishDate 2020-06-01
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).
url http://www.sciencedirect.com/science/article/pii/S2666166720300101
work_keys_str_mv AT toshihisanomura highlyefficientcrisprassociatedprotein9ribonucleoproteinbasedgenomeeditingineuglenagracilis
AT mizukiyoshikawa highlyefficientcrisprassociatedprotein9ribonucleoproteinbasedgenomeeditingineuglenagracilis
AT kengosuzuki highlyefficientcrisprassociatedprotein9ribonucleoproteinbasedgenomeeditingineuglenagracilis
AT keiichimochida highlyefficientcrisprassociatedprotein9ribonucleoproteinbasedgenomeeditingineuglenagracilis
_version_ 1724446647651401728

Similar Items