Generating gene knockout Oryzias latipes and rice field eel using TALENs method

Abstract

Transcription activator-like effector nucleases (TALENs) methods based on engineered nucleases enable precise manipulations with genomic DNA. Within the field, genetic manipulation has surpassed the proof of principle stage and is now utilized in both applied strategies and performing in economic organism. The generation and study of gene modified (GM) fish using TALENs provides a novel and essential tool for fishes molecular breeding. The medaka, Oryzias latipes, is a small, freshwater fish and an idea model organism to study reproductive mechanism. Rice Field eel is a valuable model organism of high economic importance in PR China. In this chapter, we describe microinjection of Oryzias latipes and Rice Field eel embryos, in the context of preparing, evaluating, performing gene knockout and generate gene knockout fish with TALENs in detail, which are easy to prepare and proved to be efficient in targeting of a wide range of cleavage sites. Our procedure will facilitate broader applications of TALENs in freshwater aquaculture organism.