中国比较医学杂志
中國比較醫學雜誌
중국비교의학잡지
CHINESE JOURNAL OF COMPARATIVE MEDICINE
2015年
3期
1-8
,共8页
马婧%陈炜%张旭%马元武%吕丹%高虹%张连峰
馬婧%陳煒%張旭%馬元武%呂丹%高虹%張連峰
마청%진위%장욱%마원무%려단%고홍%장련봉
Abcb1%人源化%BAC%基因敲除大鼠
Abcb1%人源化%BAC%基因敲除大鼠
Abcb1%인원화%BAC%기인고제대서
Abcb1%Humanized%BAC%Gene knock out rat
目的:敲除大鼠多药耐药基因1(Abcb1),并在基因敲除大鼠的基础上建立Abcb1人源化大鼠模型,为Abcb1相关药物代谢和药物评价研究提供更接近人类的动物模型。方法利用大片段转基因技术和CRISPR/Cas9技术相结合,建立人源化大鼠模型,利用PCR,RT-PCR和Real-time PCR的方法进行鉴定及分析。结果将包含人源Abcb1启动子和cDNA的153 kb BAC片断转入到大鼠基因组,获得稳定表达h-Abcb1基因的大鼠,同时建立了Abcb1基因敲除大鼠。通过将两者杂交建立了Abcb1人源化大鼠模型。人源大鼠与大鼠内源Abcb1表达谱有明显的区别,人源化大鼠不仅表达h-Abcb1基因,在组织表达谱方面也与人类更接近。结论建立了Abcb1基因敲除大鼠和人源化大鼠模型,Abcb1人源化模型可作为Abcb1基因相关药物代谢研究更接近人类的动物模型。
目的:敲除大鼠多藥耐藥基因1(Abcb1),併在基因敲除大鼠的基礎上建立Abcb1人源化大鼠模型,為Abcb1相關藥物代謝和藥物評價研究提供更接近人類的動物模型。方法利用大片段轉基因技術和CRISPR/Cas9技術相結閤,建立人源化大鼠模型,利用PCR,RT-PCR和Real-time PCR的方法進行鑒定及分析。結果將包含人源Abcb1啟動子和cDNA的153 kb BAC片斷轉入到大鼠基因組,穫得穩定錶達h-Abcb1基因的大鼠,同時建立瞭Abcb1基因敲除大鼠。通過將兩者雜交建立瞭Abcb1人源化大鼠模型。人源大鼠與大鼠內源Abcb1錶達譜有明顯的區彆,人源化大鼠不僅錶達h-Abcb1基因,在組織錶達譜方麵也與人類更接近。結論建立瞭Abcb1基因敲除大鼠和人源化大鼠模型,Abcb1人源化模型可作為Abcb1基因相關藥物代謝研究更接近人類的動物模型。
목적:고제대서다약내약기인1(Abcb1),병재기인고제대서적기출상건립Abcb1인원화대서모형,위Abcb1상관약물대사화약물평개연구제공경접근인류적동물모형。방법이용대편단전기인기술화CRISPR/Cas9기술상결합,건립인원화대서모형,이용PCR,RT-PCR화Real-time PCR적방법진행감정급분석。결과장포함인원Abcb1계동자화cDNA적153 kb BAC편단전입도대서기인조,획득은정표체h-Abcb1기인적대서,동시건립료Abcb1기인고제대서。통과장량자잡교건립료Abcb1인원화대서모형。인원대서여대서내원Abcb1표체보유명현적구별,인원화대서불부표체h-Abcb1기인,재조직표체보방면야여인류경접근。결론건립료Abcb1기인고제대서화인원화대서모형,Abcb1인원화모형가작위Abcb1기인상관약물대사연구경접근인류적동물모형。
Objective To knock out the Abcb1 gene of rat,and establish the Abcb1 humanized rat model based on the Abcb1 knock out rat.Methods The animal model was established using BAC and CRISPR/Cas9 technology,and was analyzed by PCR, RT-PCR and real-time PCR.Results Establishing a rat model expressing human Abcb1 stably by transfer the 153 kb BAC containing human Abcb1 promoter and cDNA into rat genome, and establishing the Abcb1 knock out rat at the same time.Establishing the Abcb1 humanized model by crossing these two strains together.The expression pattern of Abcb1 in Abcb1 humanized rat is different from the wild type rat.The Abcb1 humanized model express not only the human Abcb1 gene but also has similar expression pattern as human.Conclusions The Abcb1 knock out rat and the Abcb1 humanized rat were successfully established, and this model is close to human concerning about the drug metabolism related to Abcb1.