目的 利用成簇规律间隔短回文重复序列（CRISPR）/重组CRISPR相关核酸酶9（Cas9）技术双切口法制备成对框基因2（Pax2）敲除小鼠，为探讨Pax2基因在多个系统发育的作用提供动物模型。  方法 根据Pax2基因序列设计sgRNA，设计出的sgRNA和Cas9体外转录后显微注射到C57BL/6J 小鼠的受精卵中，F0代小鼠出生后取其基因DNA测序鉴定基因型。共获得8只F0 代小鼠，使敲除成功的F0代小鼠与野生C57BL/6J 小鼠交配，获得F1代小鼠，后均采用基因成功敲除的小鼠与C57BL/6J 小鼠进行交配，可获得稳定的Pax2基因敲除小鼠。  结果 成功获得可稳定繁殖的Pax2杂合子基因敲除小鼠，其Pax2基因缺失1628 bp；组织HE染色显示，敲除小鼠的肾小球数量明显减少；Western blotting结果显示，敲除小鼠的肾皮质Pax2蛋白表达较野生型小鼠减少。  结论 利用CRISPR/Cas9技术可成功构建Pax2杂合子基因敲除小鼠，为进一步研究Pax2基因的作用奠定基础。

Objective To establish paired box gene 2(Pax2) gene knockout mice by clusterd regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9 (Cas9) technology with double nicking method  so as to provide an animal model for studying the role of Pax2 gene in multiple development.  Methods The sgRNA was designed according to the Pax2 gene sequence, and the designed sgRNA and Cas9 were microinjected into the fertilized eggs of C57BL/6J mice in vitro. F0 generation mice were genetically sequenced to identify genotypes after birth, and a total of 8 F0 generation mice were obtained. The F0 mice were mated with wild C57BL/6J mice to obtain F1 mice, and the mice successfully knocked out were successfully mated with C57BL/6J mice to obtain stable Pax2 knockout mice.   Results The stably propagated Pax2 heterozygous knockout mice were successfully obtained, and the Pax2 gene was deleted by 1628 bp. HE staining showed that the number of glomeruli in knockout mice was significantly reduced. Western blotting result  showed that the expression of Pax2 protein in renal cortex of knockout mice was reduced compared with that in wild type mice.  Conclusion The Pax2 heterozygous knockout mice can be successfully constructed by using CRISPR/Cas9 technology, which lays a foundation for further study of the role of Pax2 gene.