| abstract |
The invention discloses a method for quickly obtaining a non-chimeric double-allele knockout animal model, which comprises the steps of firstly selecting a targeting site aiming at a gene to be knocked out, and designing and synthesizing sgRNA; then processing the male experimental animal to obtain sperms, and preparing a sperm suspension; treating a female laboratory animal to obtain oocytes and maintain fertilization activity; and finally, configuring a microinjection system by the sperm suspension, the Cas9 protein and the sgRNA, microinjecting the mixture into the oocyte, incubating to obtain an F0 generation animal, and screening to obtain a non-chimeric double-allele knockout animal model. The method is simple to operate, consumes very short time, can directly obtain the non-chimeric F0 generation in one step, has strong applicability, lays a foundation for obtaining the knockout animal model efficiently in a time-saving manner, and effectively improves the efficiency of experiments and researches. |