Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single-cell mutation detection to mutant plant regeneration | |
Choun-Sea Lin1; Chen-Tran Hsu1; Ling-Hung Yang1; Lan-Ying Lee2; Jin-Yuan Fu1; Qiao-Wei Cheng1; Fu-Hui Wu1; Han C.-W. Hsiao3; Yesheng Zhang4; Ru Zhang4 | |
刊名 | Plant Biotechnology Journal
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2018 | |
期号 | 16页码:1295–1310 |
关键词 | Protoplast Isolation Crispr/cas9 Protoplast Regeneration Single-cell Analysis |
DOI | 10.1111/pbi.12870 |
英文摘要 | Plant protoplasts are useful for assessing the efficiency of clustered regularly interspaced shortpalindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) mutagenesis. We improved theprocess of protoplast isolation and transfection of several plant species. We also developed amethod to isolate and regenerate single mutagenized Nicotianna tabacum protoplasts intomature plants. Following transfection of protoplasts with constructs encoding Cas9 and sgRNAs,target gene DNA could be amplified for further analysis to determine mutagenesis efficiency. Weinvestigated N. tabacum protoplasts and derived regenerated plants for targeted mutagenesis ofthe phytoene desaturase (NtPDS) gene. Genotyping of albino regenerants indicated that all fourNtPDS alleles were mutated in amphidiploid tobacco, and no Cas9 DNA could be detected inmost regenerated plants. |
语种 | 英语 |
内容类型 | 期刊论文 |
源URL | [http://159.226.149.26:8080/handle/152453/12295] ![]() |
专题 | 昆明动物研究所_基因起源组 |
通讯作者 | Choun-Sea Lin; Stanton B. Gelvin; Ming-Che Shih |
作者单位 | 1.Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan 2.Department of Biological Sciences, Purdue University, West Lafayette, IN, USA 3.Department of Bioinformatics and Medical Engineering, Asia University, Taichung City, Taiwan 4.State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, China 5.Institute of Life Science, National Kaohsiung Normal University, Kaohsiung, Taiwan |
推荐引用方式 GB/T 7714 | Choun-Sea Lin,Chen-Tran Hsu,Ling-Hung Yang,et al. Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single-cell mutation detection to mutant plant regeneration[J]. Plant Biotechnology Journal,2018(16):1295–1310. |
APA | Choun-Sea Lin.,Chen-Tran Hsu.,Ling-Hung Yang.,Lan-Ying Lee.,Jin-Yuan Fu.,...&Ming-Che Shih.(2018).Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single-cell mutation detection to mutant plant regeneration.Plant Biotechnology Journal(16),1295–1310. |
MLA | Choun-Sea Lin,et al."Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single-cell mutation detection to mutant plant regeneration".Plant Biotechnology Journal .16(2018):1295–1310. |
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