Study On Edition Of MSTN Gene In Rex Rabbit By CRISPR/Cas9

Since CRISPR/Cas9 is simple,low cost and high effectively,it has been developed as a genetic modification technique in recent years.Sperm-mediated gene transfer(SMGT)is a simple and efficient method for producing transgenic animals.The aim of this study was to combine SMGT with CRISPR/Cas9 to edit the MSTN gene of Rex rabbits.In this study,two sgRNA targeted on coding regions of MSTN gene were designed.Molecular cloning technology was used to construct MSTN gene knockout vector.Then the MSTN gene knockout vector and sperm were mixed,and then the MSTN gene of Rex rabbits was edited by artificial fertilization.The results showed that two MSTN knockout vectors were successfully constructed and no absence of important elements in the carrier.32 female rabbits were inseminated by artificial insemination in this study.A total of 11 female rabbits were conceived,and three of them were died.47 pups rabbits were produced in the study,of which 10 pups rabbits died.None of these rabbits found gene-edited Rex Rabbits.In this experiment,the way combine SMGT with CRISPR/Cas9 to edit the gene of Rex rabbits is immature and needs further improvement.