| 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
|
| リソース情報 |
| 藻類 |
NIES-2145 |
