CRISPR/Cas9-mediated deletion of EcMIH shortens metamorphosis time from mysis larva to postlarva of Exopalaemon carinicauda

doi:10.1016/j.fsi.2018.04.002

CORC > 海洋研究所 > 中国科学院海洋研究所 > 实验海洋生物学重点实验室

	CRISPR/Cas9-mediated deletion of EcMIH shortens metamorphosis time from mysis larva to postlarva of Exopalaemon carinicauda
	Zhang, Jiquan 1,2,3; Song, Fengge2,4 ; Sun, Yuying 1; Yu, Kuijie 2,3; Xiang, Jianhai2,3
刊名	FISH & SHELLFISH IMMUNOLOGY
	2018-06-01
卷号	77 页码:244-251
关键词	Exopalaemon carinicauda CRISPR/Cas9 Molt-inhibiting hormone Knock-out
ISSN号	1050-4648
DOI	10.1016/j.fsi.2018.04.002
通讯作者	Sun, Yuying(sunyuying125@163.com)
英文摘要	The recently emerged CRISPR/Cas9 technology is the most flexible means to produce targeted mutations at the genomic loci in a variety of organisms. In Crustaceans, molt-inhibiting hormone (MIH) is an important negative-regulatory factor and plays a key role in suppressing the molting process. However, whether precise disruption of MIH in crustacean can be achieved and successfully used to improve the development and growth has not been proved. In this research, the complementary DNA (cDNA) and genomic DNA, including flanking regions of the MIH gene (EcMIH) of ridgetail white prawn Exopalaemon carinicauda, were cloned and sequenced. Sequence analysis revealed that EcMIH was composed of three exons and two introns. Analysis by RT-PCR showed that EcMIH mainly expressed in eyestalks. During different development periods, EcMIH was highest in juvenile stage and extremely low in others but adult prawns eyestalks. In addition, we applied CRISPR/Cas9 technology to generate EcMIH knock-out (KO) prawns and then analyzed the changes in their phenotypes. We efficiently generated 12 EcMIH-KO prawns out of 250 injected one-cell stage embryos and the mutant rate reached 4.8% after embryo injection with one sgRNA targeting the second exon of EcMIH. The EcMIH-KO prawns exhibited increased the body length and shortened the metamorphosis time of larvae from mysis larva to postlarva. Meanwhile, EcMIH-KO did not cause the health problems such as early stage death or deformity. In conclusion, we successfully obtained EcMIH gene and generated EcMIH-KO prawns using CRISPR/Cas9 technology. This study will certainly lead to a wide application prospect of MIH gene in prawns breeding.
资助项目	National Natural Science Foundation of China[31172449] ; National Natural Science Foundation of China[41376165] ; National High Technology Research and Development Program of China[2012AA10A401]
WOS研究方向	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
语种	英语
出版者	ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
WOS记录号	WOS:000432644300028
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/159208]
专题	海洋研究所_实验海洋生物学重点实验室
通讯作者	Sun, Yuying
作者单位	1.Hebei Univ, Coll Life Sci, Baoding 071002, Hebei, Peoples R China 2.Chinese Acad Sci, Inst Oceanol, CAS Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 3.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266000, Peoples R China 4.Univ Chinese Acad Sci, Beijing 100039, Peoples R China
推荐引用方式 GB/T 7714	Zhang, Jiquan,Song, Fengge,Sun, Yuying,et al. CRISPR/Cas9-mediated deletion of EcMIH shortens metamorphosis time from mysis larva to postlarva of Exopalaemon carinicauda[J]. FISH & SHELLFISH IMMUNOLOGY,2018,77:244-251.
APA	Zhang, Jiquan,Song, Fengge,Sun, Yuying,Yu, Kuijie,&Xiang, Jianhai.(2018).CRISPR/Cas9-mediated deletion of EcMIH shortens metamorphosis time from mysis larva to postlarva of Exopalaemon carinicauda.FISH & SHELLFISH IMMUNOLOGY,77,244-251.
MLA	Zhang, Jiquan,et al."CRISPR/Cas9-mediated deletion of EcMIH shortens metamorphosis time from mysis larva to postlarva of Exopalaemon carinicauda".FISH & SHELLFISH IMMUNOLOGY 77(2018):244-251.