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CRISPR/Cas9-mediated deletion of EcMIH shortens metamorphosis time from mysis larva to postlarva of Exopalaemon carinicauda
Zhang, Jiquan1,2,3; Song, Fengge2,4; Sun, Yuying1; Yu, Kuijie2,3; Xiang, Jianhai2,3
2018-06-01
Source PublicationFISH & SHELLFISH IMMUNOLOGY
ISSN1050-4648
Volume77Pages:244-251
Corresponding AuthorSun, Yuying(sunyuying125@163.com)
AbstractThe 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.
KeywordExopalaemon carinicauda CRISPR/Cas9 Molt-inhibiting hormone Knock-out
DOI10.1016/j.fsi.2018.04.002
Indexed BySCI
Language英语
Funding ProjectNational Natural Science Foundation of China[31172449] ; National Natural Science Foundation of China[41376165] ; National High Technology Research and Development Program of China[2012AA10A401]
WOS Research AreaFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS SubjectFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS IDWOS:000432644300028
PublisherACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
Citation statistics
Cited Times:3[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/159209
Collection实验海洋生物学重点实验室
Corresponding AuthorSun, Yuying
Affiliation1.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
First Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Recommended Citation
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.
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