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강범창
유전체 교정 연구단
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Guidelines for C to T base editing in plants: base-editing window, guide RNA length, and efficient promoter

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Title
Guidelines for C to T base editing in plants: base-editing window, guide RNA length, and efficient promoter
Author(s)
Beum‑Chang Kang; Je Wook Woo; Sang‑Tae Kim; Su‑Ji Bae; Minkyung Choi; Jin‑Soo Kim; Sang‑Gyu Kim
Publication Date
2019-10
Journal
PLANT BIOTECHNOLOGY REPORTS, v.13, no.5, pp.533 - 541
Publisher
SPRINGER
Abstract
The Cas9 protein fused with a cytidine deaminase can induce C to T substitutions at a specific site when directed by a guide RNA. Here, we compared the substitution activity and the substitution range of two base-editing systems, APOBEC1-nCas9 and nCas9-PmCDA1, in the protoplasts of Glycine max, Brassica napus, and Nicotiana tabacum. To prevent unwanted nucleotide substitution, we manipulated the length of guide RNA and found the change of nucleotide substitution activity in the target window of nCAS9-PmCDA1. Based on these results, the specific C to T conversion in the acetolactate synthase gene of N. tabacum was induced to generate herbicide-resistant plants. During the screening of herbicide-resistant plants, we found that ubiquitin promoter-driven base-editor system was much efficient than 35S promoter-driven base-editor system. This study provides guidelines on which a base editor to use and describes how to fine-tune a guide RNA for precise substitutions in plants. © Korean Society for Plant Biotechnology 2019
URI
https://pr.ibs.re.kr/handle/8788114/6836
ISSN
1863-5466
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > Journal Papers (저널논문)
Files in This Item:
1910_강범창_Plant Biotechnology Reports.pdfDownload

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