The invention relates to the technical field of targeted editing of a buffalo 18S rDNA
gene, in particular to a method for targeted editing of the buffalo 18S rDNA
gene by virtue of an adenovirus
system. According to the method provided by the invention, an autonomously constructed 18S
gene editing vector is used for editing the buffalo 18S rDNA gene; a constructed adenovirus vector consists of Cas9
protein and sgRNA; and then, the vector is subjected to adenovirus packaging. The buffalo 18S rDNA gene is edited on the basis of the accurate and efficient performances of a Cas9 editing gene, a strong
bearing capacity of adenovirus on exogenous genes as well as a strong infection capacity of the adenovirus on primary cells, so that a
bottleneck that the buffalo primary cells are difficult for
transfection; and the packaged adenovirus can efficiently infect buffalo primary mammary epithelial cells and can efficiently edit the buffalo 18S rDNA gene.