論文 - 詳細
| RRC ID | 89447 |
|---|---|
| 著者 | Moroi K, Yamamoto T, Kurita T. |
| タイトル | Double-strand break-free and transgene-free genome editing in the microalga Nannochloropsis oceanica using removable vectors containing the CRISPR base editing system. |
| ジャーナル | Sci Rep |
| Abstract |
The accumulation of lipids by algae makes them attractive for carbon-neutral fuel production; however, the industrial-scale production of algal lipids has yet to be achieved. Currently, researchers are trying to improve the lipid productivity of algal strains using genome editing for molecular breeding with CRISPR-Cas9, which allows the efficient alteration of genomic information. However, CRISPR-based gene modification via double-strand breaks sometimes induces unintended large deletions that are toxic to host cells. Here, we applied the cytidine base editor combined with an episomal vector backbone containing a centromere and autonomous replication sequence to the microalga Nannochloropsis oceanica. The cytosine base editor introduces cytidine-to-thymidine base substitutions using deaminase without double-strand breaks, and an episomal vector enables plasmid removal after base substitution. We succeeded in inducing cytidine-to-thymidine substitution at the six target sites of five endogenous genes. The base substitution activity ranged from 29.2% to 47.6% on cytidine bases at the 16th to 19th positions from the protospacer adjacent motifs. The removal of base editor plasmids was also detected, which is essential for constructing transgene-free strains. Our results provide insights into the applicability of further technologies in the genetic modification of microalgae. |
| 巻・号 | 15(1) |
| ページ | 42431 |
| 公開日 | 2025-11-27 |
| DOI | 10.1038/s41598-025-26657-y |
| PII | 10.1038/s41598-025-26657-y |
| PMID | 41309901 |
| PMC | PMC12661010 |
| MeSH | CRISPR-Cas Systems* DNA Breaks, Double-Stranded Gene Editing* / methods Genetic Vectors* / genetics Microalgae* / genetics Plasmids / genetics Stramenopiles* / genetics Transgenes |
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オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 2 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 藻類 | NIES-2145 |