Species: Nile tilapia (Oreochromis niloticus)

Country of 1st Author: United Kingdom
Year: 2020

Institute	University of Stirling Stirling UK
Editing Method	Embryo edit
SDN Type	SDN-1
Delivery	Injecting Cas9 mRNA and sgRNAs into Nile tilapia embryos at 1 cell stage.
Foreign DNA	No
Undergone Regulatory Process	No

Target Gene(s)

Gene Name	Gene Trait	Trait Category	NGT	Methods/Remarks
piwil2 Piwi Like RNA-Mediated Gene Silencing 2		Reproduction	CRISPR Cas9 (spCas9)	Two sgRNAs were designed using CRISPR RGEN tool (https://www.rgenome.net/) and Benchling (https://benchling.com) and the potential off-target sites were checked by in silico off-target analysis using NCBI BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi) and Cas-OFFinder (https://www.rgenome.net/cas-offinder/). Any sgRNAs containing potential off-targets which had more than 12 nt identical to the seed sequences adjacent to the protospacer adjacent motif (PAM), out of the 20 nt sgRNAs, were excluded

References, Documents, and Links

• Physiological impact and comparison of mutant screening methods in piwil2 KO founder Nile tilapia produced by CRISPR/Cas9 system