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

Toshihisa Nomura; Mizuki Yoshikawa; Kengo Suzuki; Keiichi Mochida

doi:10.1016/j.xpro.2020.100023

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

Toshihisa Nomura, Mizuki Yoshikawa, Kengo Suzuki, Keiichi Mochida

Research output: Contribution to journal › Article › peer-review

8 Citations (Scopus)

Abstract

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).

Original language	English
Article number	100023
Journal	STAR Protocols
Volume	1
Issue number	1
DOIs	https://doi.org/10.1016/j.xpro.2020.100023
Publication status	Published - Jun 19 2020

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Neuroscience(all)
Immunology and Microbiology(all)

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10.1016/j.xpro.2020.100023

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title = "Highly Efficient CRISPR-Associated Protein 9 Ribonucleoprotein-Based Genome Editing in Euglena gracilis",

abstract = "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).",

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AU - Mochida, Keiichi

N1 - Funding Information: We thank Aya Ide and Hiromi Ojima for their support in the acquisition of the photos in this experimental protocol. Methodology, T.N.; Investigation, T.N. and M.Y.; Resources, K.S. Writing ? Original Draft, T.N. and K.M. K.S. is an employee and shareholder of euglena Co. Ltd. T.N. K.S. and K.M. have applied for a patent for this method. Publisher Copyright: © 2020 The Author(s)

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N2 - 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).

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

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ASJC Scopus subject areas

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