大鼠品系PGR-iCre构建

Construction of rat strain PGR-iCre

采用CRISPR/Cas9基因编辑技术首次制备用于子宫基因敲除的PGR-iCre大鼠。PGR-iCre大鼠与含有Flox报告基因Tdtomato大鼠(Tdtomatof/f)杂交得到PGRCre+/-- Tdtomatof/+大鼠。利用real-time PCR、免疫组化、荧光检测等技术检测Tdtomato基因在PGRCre+/-- Tdtomatof/+大鼠不同器官表达,间接检测PGR驱动的Cre重组酶组织特异性表达。结果表明,PGR驱动Cre重组酶在大鼠子宫、卵巢、输卵管、乳腺、下丘脑、垂体、脾脏、肾脏、肺等器官不同程度表达;未检测到Cre重组酶在心脏、肝脏、肌肉中表达。综上,PGR-iCre大鼠可作为子宫基因敲除工具鼠研究大鼠子宫表达基因的相关功能。

CRISP/Cas9 genome editing technology was used to generate a new iCre knock-in rat strain for the first time. PGR- iCre rats were crossed with a rat with a floxed reporter gene Tdtomato (Tdtomatof/f) to obtain PGRCre +/--Tdtomatof/+ rats. The tissue-specific expression of Cre recombinase was indicated indirectly by detecting the expression of Tdtomato gene in different organs of PGRCre +/-- Tdtomatof/+ rats by real-time PCR, immunohistochemistry and immunofluorescence. The results showed that the PGR- driven Cre recombinase could express in rat uterus, ovary, fallopian tube, breast, hypothalamus, pituitary, spleen, kidney and lung in different degrees;no expression of Cre recombinase was detected in heart, liver and muscle. Taken together, these newly established PGR-iCre rats would facilitate the study of gene functions expressed in the rat uterus.