Production of germ cell-less by the dead end gene knockout in rainbow trout (Oncorhynchus mykiss) embryos using CRISPR/Cas system

• Grant number: 15/24148-2
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: January 28, 2016
• End date: May 27, 2016
• Field of knowledge: Agronomical Sciences - Fishery Resources and Fishery Engineering - Inland Water Fishery Resources
• Principal Investigator: Rafael Henrique Nóbrega
• Grantee: Melanie Digmayer
• Supervisor: Goro Yoshizaki
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Institution abroad: Tokyo University of Marine Science and Technology (TUMSAT), Japan
• Associated to the scholarship: 13/20450-0 - Molecular markers and cryopreservation of spermatogonial stem cells of common carp, Cyprinus carpio and zebrafish, Danio rerio, BP.PD

Abstract

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system is a simple and efficient method for targeted genome editing. Because of the ease production of targeting RNA, the CRISPR/ Cas9 system is a promising tool for genome engineering in several areas, including aquaculture. Surrogate valuable fish broodstock has been achieved by allogeneic and xenogeneic germ cell transplantation. Trout-derived offspring from salmon were obtained when trout primordial germ cell (PGC) in salmon are injected into newly-hatched salmon recipients. However the efficiency of this technique is low due to the competition between donor and endogenous PGC from available niches. The use of germ cell-less recipients is an alternative option to increase the efficiency of the transplantation. The dead end (dnd) is a gene specifically expressed in PGCs and has been shown to be essential PGC migration, maintenance and survival during embryogenesis. In this current project, we aim to generate trout germ cell-less recipients by using CRISPR/Cas9 for dnd. If generated, this suitable model will increase transplantation efficiency. (AU)