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Kang, Beum-Chang
유전체 교정 연구단
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Chloroplast and mitochondrial DNA editing in plants

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Title
Chloroplast and mitochondrial DNA editing in plants
Author(s)
Beum-Chang Kang; Su-Ji Bae; Seonghyun Lee; Jeong Sun Lee; Annie Kim; Hyunji Lee; Gayoung Baek; Huiyun Seo; Jihun Kim; Jin-Soo Kim
Publication Date
2021-07
Journal
NATURE PLANTS, v.7, no.7, pp.899 - 905
Publisher
NATURE RESEARCH
Abstract
Plant organelles including mitochondria and chloroplasts contain their own genomes, which encode many genes essential for respiration and photosynthesis, respectively. Gene editing in plant organelles, an unmet need for plant genetics and biotechnology, has been hampered by the lack of appropriate tools for targeting DNA in these organelles. In this study, we developed a Golden Gate cloning system(1), composed of 16 expression plasmids (8 for the delivery of the resulting protein to mitochondria and the other 8 for delivery to chloroplasts) and 424 transcription activator-like effector subarray plasmids, to assemble DddA-derived cytosine base editor (DdCBE)(2) plasmids and used the resulting DdCBEs to efficiently promote point mutagenesis in mitochondria and chloroplasts. Our DdCBEs induced base editing in lettuce or rapeseed calli at frequencies of up to 25% (mitochondria) and 38% (chloroplasts). We also showed DNA-free base editing in chloroplasts by delivering DdCBE mRNA to lettuce protoplasts to avoid off-target mutations caused by DdCBE-encoding plasmids. Furthermore, we generated lettuce calli and plantlets with edit frequencies of up to 99%, which were resistant to streptomycin or spectinomycin, by introducing a point mutation in the chloroplast 16S rRNA gene.
URI
https://pr.ibs.re.kr/handle/8788114/10733
DOI
10.1038/s41477-021-00943-9
ISSN
2055-026X
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > 1. Journal Papers (저널논문)
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