CRISPR/Cas9-mediated mutagenesis reveals the roles of calaxin in gastropod larval cilia
文献类型：期刊论文
| 作者 | Huan, Pin1,2,3,4; Cui, Menglu1,2,4; Wang, Qian1,2,4 ; Liu, Baozhong1,2,3,4
|
| 刊名 | GENE
 |
| 出版日期 | 2021-06-30 |
| 卷号 | 787页码:10 |
| ISSN号 | 0378-1119 |
| 关键词 | Calaxin CRISPR Cas9 Gene editing Mollusk |
| DOI | 10.1016/j.gene.2021.145640 |
| 通讯作者 | Liu, Baozhong(bzliu@qdio.ac.cn) |
| 英文摘要 | Obtaining detectable knockout phenotypes in the G0 generation is essential for gene function studies. Although CRISPR/Cas9-mediated gene editing has been employed to knock out molluscan genes, detectable phenotypes in the G0 generation have not been reported in these animals. In this study, we determined the knockout phenotype of a cilium-related gene, calaxin, using CRISPR/Cas9 technology in the gastropod mollusk Lottia goshimai. Injections with the Cas9-sgRNA complex caused approximately 30?80% of the injected larvae to exhibit a shortcilia phenotype characteristic of shortened cilia and decreased motility in the larvae. This phenotype was detectable in the G0 generation and was consistent for two independent sgRNAs. Genotyping of the injected larvae revealed various types of deletions and insertions in the target gene, which occurred in all sequences from the short-cilia larvae. This result indicated that the short-cilia phenotype was indeed caused by calaxin knockout. This possibility was supported by an RNAi assay targeting calaxin, which produced a highly similar short-cilia phenotype. We observed that a single SNP in the target sequences of the sgRNAs could show varied effects on the efficiency of mutagenesis. These results help to establish a foundation for future studies on molluscan gene editing using the CRISPR/Cas9 technique and contribute to the body of knowledge on molluscan ciliary functions. |
| 资助项目 | National Key R&D Program of China[2018YFD0900104] ; China Agriculture Research System[CARS-49] ; National Natural Science Foundation of China[41776157] ; Marine S&T Fund of Shandong Province for Pilot National Laboratory for Marine Science and Technology (Qingdao)[2018SDKJ0302-1] |
| WOS研究方向 | Genetics & Heredity |
| 语种 | 英语 |
| 出版者 | ELSEVIER |
| WOS记录号 | WOS:000642475500001 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/170594]  |
| 专题 | 海洋研究所_实验海洋生物学重点实验室 |
| 通讯作者 | Liu, Baozhong |
| 作者单位 | 1.Univ Chinese Acad Sci, Beijing 100039, Peoples R China 2.Chinese Acad Sci, Ctr Ocean Megasci, 7 Nanhai Rd, Qingdao 266071, Peoples R China 3.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266000, Peoples R China 4.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, 7 Nanhai Rd, Qingdao 266071, Peoples R China |
| 推荐引用方式 GB/T 7714 | Huan, Pin,Cui, Menglu,Wang, Qian,et al. CRISPR/Cas9-mediated mutagenesis reveals the roles of calaxin in gastropod larval cilia[J]. GENE,2021,787:10. |
| APA | Huan, Pin,Cui, Menglu,Wang, Qian,&Liu, Baozhong.(2021).CRISPR/Cas9-mediated mutagenesis reveals the roles of calaxin in gastropod larval cilia.GENE,787,10. |
| MLA | Huan, Pin,et al."CRISPR/Cas9-mediated mutagenesis reveals the roles of calaxin in gastropod larval cilia".GENE 787(2021):10. |
入库方式： OAI收割
来源：海洋研究所
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