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CRISPR/Cas9-mediated mutagenesis reveals the roles of calaxin in gastropod larval cilia
Huan, Pin1,2,3,4; Cui, Menglu1,3,4; Wang, Qian1,3,4; Liu, Baozhong1,2,3,4
2021-06-30
Source PublicationGENE
ISSN0378-1119
Volume787Pages:10
Corresponding AuthorLiu, Baozhong(bzliu@qdio.ac.cn)
AbstractObtaining 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.
KeywordCalaxin CRISPR Cas9 Gene editing Mollusk
DOI10.1016/j.gene.2021.145640
Indexed BySCI
Language英语
Funding ProjectNational 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 Research AreaGenetics & Heredity
WOS SubjectGenetics & Heredity
WOS IDWOS:000642475500001
PublisherELSEVIER
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Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/170595
Collection实验海洋生物学重点实验室
Corresponding AuthorLiu, Baozhong
Affiliation1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, 7 Nanhai Rd, Qingdao 266071, Peoples R China
2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266000, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100039, Peoples R China
4.Chinese Acad Sci, Ctr Ocean Megasci, 7 Nanhai Rd, Qingdao 266071, Peoples R China
First Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences;  Center for Ocean Mega-Science, Chinese Academy of Sciences
Corresponding Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences;  Center for Ocean Mega-Science, Chinese Academy of Sciences
Recommended Citation
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.
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