CRISPR/Cas9-mediated mutagenesis reveals the roles of calaxin in gastropod larval cilia

doi:10.1016/j.gene.2021.145640

IOCAS-IR > 实验海洋生物学重点实验室

	CRISPR/Cas9-mediated mutagenesis reveals the roles of calaxin in gastropod larval cilia
	Huan, Pin1,2,3,4 ; Cui, Menglu 1,3,4; Wang, Qian1,3,4 ; Liu, Baozhong1,2,3,4
	2021-06-30
发表期刊	GENE
ISSN	0378-1119
卷号	787 页码:10
通讯作者	Liu, Baozhong([email protected])
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.
关键词	Calaxin CRISPR Cas9 Gene editing Mollusk
DOI	10.1016/j.gene.2021.145640
收录类别	SCI
语种	英语
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
WOS类目	Genetics & Heredity
WOS记录号	WOS:000642475500001
出版者	ELSEVIER
引用统计	被引频次：7[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/170595
专题	实验海洋生物学重点实验室
通讯作者	Liu, Baozhong
作者单位	1.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
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.

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