NgAgo-based fabp11a gene knockdown causes eye developmental defects in zebrafish

A recent report of genome editing using Natronobacterium gregoryi Argonaute (NgAgo) with guide DNA (gDNA) in human cells prompted us to explore the utility of this protein for in vivo genetic manipulation in zebrafish (Danio rerio). Zebrafish is a model organism that offers several distinct advantages for studying genetics, developmental biology, vascular biology and disease modeling. In the last several years, loss-of-function genomic editing techniques, including zinc-finger nucleases (ZFNs), artificial transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) 9 system (CRISPR/Cas9), have been adopted for zebrafish. Recently the gDNA/NgAgo system has elicited much interest because of some unique advantages: low tolerance to guide-target mismatch, minimum off-target effects, and easy to design. Here we investigated whether the gDNA/NgAgo system could be used to manipulate zebrafish genes in vivo using fabp11a as a test case.