Use of CRISPR/Cas9 technology efficiently targetted goat myostatin through zygotes microinjection resulting in double-muscled phenotype in goats

Introduction Myostatin gene (MSTN) can inhibit the proliferation of myoblast, which in turn promotes muscle growth and inhibits adipocyte differentiation in livestock. MSTN mutation may lead to muscle hypertrophy or double-muscled (DM) phenotype. MSTN mutation animal, such as sheep, dog and rabbit have generated through CRISPR/Cas9 technology. However, goats with promising MSTN mutation have not been generated. Methods We designed two sgRNAs loci targeting exon3 of MSTN gene to destroy the MSTN cysteines knots. We got 7 goats from 7 recipients, in which 6 were MSTN KO goats, with a mutation-rate is of 85.7%. Results Destroyed cysteine knots caused MSTN structure inactivation. The average body weight gain per day of MSTN KO goats was significant higher than that of WT goats. Discussion MSTN KO goats showed abnormal sugar, fat and protein metabolism compared with wild-type controls (MSTN +/+ ). Inheritance of mutations was observed in offspring of MSTN KO goats by PCR analysis.