지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2024.6
- 수록면
- 415 - 424 (10page)
- DOI
- 10.1007/s11816-024-00901-9
이용수
초록· 키워드
오류제보하기
The CRISPR/Cas9-based gene editing system for the direct delivery of pre-assembled Cas9 ribonucleoproteins (RNPs), consisting of a Cas9 nuclease and a single guide RNA (sgRNA), into plant protoplasts enables DNA-free gene editing with- out introducing foreign gene into plants. Here, we described the optimization of CRISPR/Cas9 RNPs delivery into cabbage protoplasts for efficient DNA-free gene editing. We determined the insertion and deletion (indel) frequency of BoMYBL2-1, a negative regulatory gene for anthocyanin biosynthesis in cabbage (Brassica oleracea var. capitata). We optimized the molar ratio of Cas9 to sgRNA and the incubation time of RNP–protoplast transfection to enhance the indel frequency under various conditions. Based on the BoMYBL2-1 nucleotide sequences, we designed nine sgRNAs to target BoMYBL2-1. Our in vitro digestion assay showed that all sgRNAs were able to cleave the targeted fragment. When the sgRNA and Cas9 proteins were subsequently transfected into protoplasts isolated from cabbage cotyledons, the deep sequencing results showed that the indel frequency of sgRNAs in BoMYBL2-1 was the highest (7.4%) with sgRNA3. We compared various molar ratios of Cas9 and sgRNA and incubation times of RNP–protoplast transfection to optimize transfection and ensure high indel frequency. The highest frequency was observed when the Cas9:sgRNA ratio was 1:10. Furthermore, when the incubation time for RNP–protoplast transfection was 1 min and 3 min, the indel frequency was higher than 25%. Altogether, these results provide valuable information on the optimized conditions for high-efficiency gene editing using CRISPR/Cas9 RNP delivery into cabbage protoplasts.
목차
등록된 정보가 없습니다.
참고문헌 (0)
참고문헌 신청함께 읽어보면 좋을 논문
논문 유사도에 따라 DBpia 가 추천하는 논문입니다. 함께 보면 좋을 연관 논문을 확인해보세요!
-
DNA-free Genome Editing using Efficient Protoplast Culture and CRISPR/Cas9 Ribonucleoprotein Delivery in Tomato
한국원예학회 학술발표요지
2023 .05
-
DNA-free Genome Editing of the Ethylene Biosynthesis Gene in Carnation Protoplasts using CRISPR-Cas9 Ribonucleoprotein Complexes
한국원예학회 학술발표요지
2023 .05
-
Optimization of an Efficient Protoplast Regeneration System for Genome Editing in Chinese Cabbage
한국원예학회 학술발표요지
2023 .10
-
DNA‑free mutagenesis of GIGANTEA in Brassica oleracea var. capitata using CRISPR/Cas9 ribonucleoprotein complexes
Plant Biotechnology Reports
2019 .01
-
Efficient genome editing using CRISPR?Cas9 RNP delivery into cabbage protoplasts via electro-transfection
Plant Biotechnology Reports
2020 .01
-
Development of Radish Variety with Less Spicy Taste and High Glucoraphanin using CRISPR/Cas9 RNP Technology
한국원예학회 학술발표요지
2021 .05
-
벼에서 CRISPR/Cas9 활용 고빈도 유전자 편집 방법
Journal of Plant Biotechnology
2017 .01
-
현사시나무 원형질체에서 리보핵산단백질을 활용한 유전자 교정 방법 연구
Journal of Plant Biotechnology
2021 .01
-
Selection of Candidate Genes for Developing Drought-Tolerant Plants Using CRISPR/Cas9-Mediated Gene Editing in Chinese Cabbage (Brassica rapa)
한국원예학회 학술발표요지
2024 .11
-
CRISPR/Cas9-mediated Gene Editing of GIGANTE Enhances Abiotic Stress Tolerance in Chinese cabbages
한국원예학회 학술발표요지
2020 .11
-
CRISPR–Cas13d in plant biology: an insight
Plant Biotechnology Reports
2024 .06
-
Overview of CRISPR/Cas9: a chronicle of the CRISPR system and application to ornamental crops
Korean Journal of Agricultural Science
2020 .12
-
Generation of Late-flowering Radish (Raphanus sativus) using CRISPR/Cas9 RNP Technology
한국원예학회 학술발표요지
2021 .05
-
CRISPR/Cas9-mediated Editing of Genes to Reduces Tomato Fruit Allergen
한국원예학회 학술발표요지
2023 .05
-
Production of Male Sterile Chinese Cabbage Using CRISPR/Cas9 System
한국원예학회 학술발표요지
2022 .11
-
벼의 CRISPR/Cas9 기반 유전자 교정을 통한 내병성 및 내재해성 증진 연구 동향
한국육종학회지
2022 .12
-
Enhancing Black-rot Resistance in Cabbage using CRISPR/Cas9 System
한국원예학회 학술발표요지
2023 .10
-
Induction of Cabbage Mutation with Delayed Bolting using a CRISPR-cas9 System
한국원예학회 학술발표요지
2022 .05
-
Development of Transgenic Chinese Cabbage Using CRISPR/Cas9 to Control Self-incompatibility
한국원예학회 학술발표요지
2018 .10
-
Development of Drought Tolerance Transgenic Chinese Cabbage Using CRISPR-Cas9 System
한국원예학회 학술발표요지
2018 .10
최근 본 자료
댓글(0)
0