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Genome editing using a DNA-free clustered regularly interspaced short palindromic repeats-Cas9 system in green seaweed Ulva prolifera
| Title: | Genome editing using a DNA-free clustered regularly interspaced short palindromic repeats-Cas9 system in green seaweed Ulva prolifera |
| Authors: | Ichihara, Kensuke Browse this author →KAKEN DB | | Yamazaki, Tomokazu Browse this author | | Kawano, Shigeyuki Browse this author →KAKEN DB |
| Keywords: | 2-fluoroadenine | | adenine phosphoribosyl transferase | | green alga | | reverse genetics | | ribonucleoprotein complex | | transfection |
| Issue Date: | 3-Nov-2021 |
| Publisher: | John Wiley & Sons |
| Journal Title: | Phycological research |
| Volume: | 70 |
| Issue: | 1 |
| Start Page: | 50 |
| End Page: | 56 |
| Publisher DOI: | 10.1111/pre.12472 |
| Abstract: | Although the green seaweed Ulva is one of the most common seaweeds in the coastal regions with well-studied ecological characteristics, few reverse genetic technologies have been developed for it. The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system is a simple genome-editing technology based on a ribonucleoprotein (RNP) complex composed of an endonuclease and programmable RNA to target particular DNA sequences. Genome editing makes it possible to generate mutations on a target gene in non-model organisms without established transgenic technologies. In this study, we applied the CRISPR-Cas9 RNP genome-editing system to the green seaweed Ulva prolifera, using polyethylene glycol (PEG)-mediated transfection. Our experimental system disrupts a single gene (UpAPT) encoding adenine phosphoribosyl transferase (APT) and generates a resistant phenotype for gametophytes cultured in a medium with toxic compound 2-fluoroadenine. The PEG-mediated transfection used for gametes resulted in 2-fluoroadenine-resistant strains containing short indels or substitutions on UpAPT. Our results showed that the CRISPR-Cas9 system with PEG-mediated transfection was efficient for genome editing in Ulva. |
| Rights: | This is the peer reviewed version of the following article: Ichihara, K., Yamazaki, T. and Kawano, S. (2021), Genome editing using a DNA-free clustered regularly interspaced short palindromic repeats-Cas9 system in green seaweed Ulva prolifera. Phycological Res., which has been published in final form at https://doi.org/10.1111/pre.12472. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/87604 |
| Appears in Collections: | 北方生物圏フィールド科学センター (Field Science Center for Northern Biosphere) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 市原 健介
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