Institutional Repository of Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
Easy-to-Use CRISPR-Cas9 Genome Editing in the Cultured Pacific Abalone (Haliotis discus hannai) | |
Li, Ruohui1,2,3,4; Xu, Yue1,2,3; Wu, Fucun1,2,3,4,6,7; Peng, Zhangjie1,2,3,4; Chan, Jiulin1,2,3; Zhang, Linlin1,2,3,4,5,6,7 | |
2024-02-01 | |
发表期刊 | CRISPR JOURNAL (IF:5.343[JCR-2019],5.343[5-Year]) |
ISSN | 2573-1599 |
卷号 | 7期号:1页码:41-52 |
通讯作者 | Wu, Fucun(wufucun@qdio.ac.cn) ; Zhang, Linlin(linlinzhang@qdio.ac.cn) |
摘要 | The Pacific abalone is an important aquaculture shellfish and serves as an important model in basic biology study. However, the study of abalone is limited by lack of highly efficient and easy-to-use gene-editing tools. In this paper, we demonstrate efficient gene knockout in Pacific abalone using CRISPR-Cas9. We developed a highly effective microinjection method by nesting fertilized eggs in a low-concentration agarose gel. We identified the cilia developmental gene beta-tubulin and light-sensitive transmembrane protein r-opsin as target genes and designed highly specific sgRNAs for modifying their genomic sequences. Sanger sequencing of the genomic regions of beta-tubulin and r-opsin genes from injected larvae identified various genomic long-fragment deletions. In situ hybridization showed gene expression patterns of beta-tubulin and r-opsin were significantly altered in the mosaic mutants. Knocking out beta-tubulin in abalone embryos efficiently affected cilia development. Scanning electron microscopy and swimming behavior assay showed defecting cilia and decreased motility. Moreover, knocking out of r-opsin in abalone embryos effectively affected the expression and development of eyespots. Overall, this work developed an easy-to-use mosaic gene knockout protocol for abalone, which will allow researchers to utilize CRISPR-Cas9 approaches to study unexploited abalone biology and will lead to novel breeding methods for this aquaculture species. |
DOI | 10.1089/crispr.2023.0070 |
收录类别 | SCI |
语种 | 英语 |
资助项目 | National Key R&D Program of China[2022YFD2401400]; Strategic Priority Research Program of the Chinese Academy of Sciences[XDB42000000]; Marine S&T Fund of Shandong Province for Pilot National Laboratory for Marine Science and Technology (Qingdao)[2022QNLM030004]; Key Research and Development Program of Shandong[2022LZGC015] |
WOS研究方向 | Genetics & Heredity |
WOS类目 | Genetics & Heredity |
WOS记录号 | WOS:001162177200004 |
出版者 | MARY ANN LIEBERT, INC |
WOS关键词 | CRISPR/CAS9 ; SPERM |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | http://ir.qdio.ac.cn/handle/337002/185107 |
专题 | 实验海洋生物学重点实验室 深海极端环境与生命过程研究中心 |
通讯作者 | Wu, Fucun; Zhang, Linlin |
作者单位 | 1.Chinese Acad Sci, Inst Oceanol, CAS & Shandong Prov Key Lab Expt Marine Biol, Qingdao, Peoples R China 2.Chinese Acad Sci, Inst Oceanol, Ctr Deep Sea Res, Qingdao, Peoples R China 3.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China 4.Univ Chinese Acad Sci, Coll Marine Sci, Beijing, Peoples R China 5.Chinese Acad Sci, Key Lab Breeding Biotechnol & Sustainable Aquacult, Wuhan, Peoples R China 6.Chinese Acad Sci, Inst Oceanol, CAS & Shandong Prov Key Lab Expt Marine Biol, 7 Naihai Rd, Qingdao 266071, Peoples R China 7.Chinese Acad Sci, Inst Oceanol, Ctr Deep Sea Res, 7 Naihai Rd, Qingdao 266071, Peoples R China |
推荐引用方式 GB/T 7714 | Li, Ruohui,Xu, Yue,Wu, Fucun,et al. Easy-to-Use CRISPR-Cas9 Genome Editing in the Cultured Pacific Abalone (Haliotis discus hannai)[J]. CRISPR JOURNAL,2024,7(1):41-52. |
APA | Li, Ruohui,Xu, Yue,Wu, Fucun,Peng, Zhangjie,Chan, Jiulin,&Zhang, Linlin.(2024).Easy-to-Use CRISPR-Cas9 Genome Editing in the Cultured Pacific Abalone (Haliotis discus hannai).CRISPR JOURNAL,7(1),41-52. |
MLA | Li, Ruohui,et al."Easy-to-Use CRISPR-Cas9 Genome Editing in the Cultured Pacific Abalone (Haliotis discus hannai)".CRISPR JOURNAL 7.1(2024):41-52. |
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