Species: Loach (Paramisgurnus dabryanus)

Country of 1st Author: China
Year: 2019

Institute	Huazhong Agricultural University Wuhan, 437000, Hubei CN
Editing Method	Embryo edit
SDN Type	SDN-1
Delivery	One-cell stage fertilized eggs were fixed on an agarose gel block with size matched holes in a row. A 2 nL mix containing 50 ng/µL gRNA and 150 ng/µL Cas9 mRNA in RNase-free water was injected in every fertilized egg at the yolk/blastomere boundary through the chorion using picospritzer III microinjector (Parker Hannifin, S/N 1718) and needles produced from a microelectrode drawing apparatus (Sutter, P-2000). We were able to inject 10–20 eggs in 1 min.
Foreign DNA	No
Undergone Regulatory Process	No

Target Gene(s)

Gene Name	Gene Trait	Trait Category	NGT	Methods/Remarks
TYR Tyrosinase	Pigmentation - melanin reduction	Colour	CRISPR Cas9 (spCas9)

References, Documents, and Links

• Production of a mutant of large-scale loach Paramisgurnus dabryanus with skin pigmentation loss by genome editing with CRISPR/Cas9 system