The invention relates to a method for establishing a Gadd45a knockout rabbit model by adopting a knockout technology and belongs to the technical field of
biotechnology. The invention aims to establish a rabbit model by knocking out GADD45a
gene by utilizing a Grispr / cas9 technology and to provide the method for establishing the Gadd45a knockout rabbit model by adopting the knockout technology andfor researching the influence of the
gene on animal liver. The method provided by the invention comprises the following steps: establishing sgRNA; synthesizing double-stranded
DNA; linearizing p UC-57 carrier; linking p UC-57 carrier with double-stranded
DNA; converting; performing
monoclonal picking and
plasmid DNA extraction; identifying
plasmid sequence; expressing
plasmid with CAS9; and performing
digestion linearization for
in vitro transcription. Through related detection, the invention successfully acquires the Gadd45a knockout rabbit model; the model is established for simulating theclinic
pathological processes of the liver diseases, such as,
fatty liver, liver
cirrhosis and
liver cancer after giving the corresponding
alcohol stimulation, is capable of effectively forecasting the clinic application effects of new vaccine, new drugs and new diagnostic reagents, and meanwhile, and is capable of greatly reducing the risk in researching and developing the new drugs; and a basismodel is supplied for the
clinical research.