| 慢病毒介导的CYP3A11基因knock-down小鼠的建立 |
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| 引用本文： | 庞浩,刘昌峨,刘勤,王露露,郭科男,王勇,魏泓.慢病毒介导的CYP3A11基因knock-down小鼠的建立[J].第三军医大学学报,2010,32(5). |
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| 作者姓名： | 庞浩 刘昌峨 刘勤 王露露 郭科男 王勇 魏泓 |
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| 作者单位： | 第三军医大学基础医学部实验动物学教研室,重庆,400038 |
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| 基金项目： | 国家自然科学基金(30700076)~~ |
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| 摘 要： | 目的构建和筛选小鼠P4503A11基因miR RNAi慢病毒载体,建立CYP3A11基因knock-down小鼠。方法利用Ravi Sachidanandam's Lab的在线软件设计沉默小鼠P4503A11基因的miRNA所对应的shRNA序列,PCR扩增后克隆到PCI-GFP-MiR质粒中,其再与慢病毒载体FUW进行重组。将psPAX2和pMD2.G两个载体和FUW-GFP-MiR-shRNA重组慢病毒载体共转染293FT细胞,包装成病毒。用包装好的病毒液感染293FT细胞后,并利用GFP蛋白表达水平进行滴度测定。将上述重组慢病毒载体分别转染FVB/N小鼠肝细胞,转染48h后,检测P4503A11基因mRNA表达水平的变化。选取干扰效果最好的FUW-GFP-MiR-shRNA1重组慢病毒载体进行制备基因knock-down小鼠模型。用显微注射法将浓缩的shRNA1病毒液注射至FVB/N小鼠12细胞期胚胎透明带下。将注射后发育至22细胞期的胚胎移植至假孕受体母鼠,得F0代小鼠。在紫外光照射下利用滤光片观察GFP在小鼠活体内的表达水平。结果DNA测序结果显示3个重组慢病毒载体均与所设计的shRNA序列一致。检...
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| 关 键 词： | 细胞色素P-450 CYP3A knock-down 微RNAs 慢病毒 小鼠 |
Establishment of CYP3A11 gene knock-down mouse by lentiviral transgenesis |
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| Pang Hao,Liu Change,Liu Qin,Wang Lulu,Guo Kenan,Wang Yong,Wei Hong.Establishment of CYP3A11 gene knock-down mouse by lentiviral transgenesis[J].Acta Academiae Medicinae Militaris Tertiae,2010,32(5). |
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| Authors: | Pang Hao Liu Change Liu Qin Wang Lulu Guo Kenan Wang Yong Wei Hong |
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| Affiliation: | Department of Laboratory Animal Science;College of Basic Medical Sciences;Third Military Medical University;Chongqing;400038;China |
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| Abstract: | Objective To construct miR RNAi lentiviral vectors of mouse cytochrome P450 3A11 and establish CYP3A11 gene knock-down mouse model.Methods The corresponding shRNA sequences of miRNA for silencing the mouse P450 3A11 gene were designed by on-line designer software on Ravi Sachidanandam's Lab Website and synthesized by PCR.The PCR products were cloned into the PCI-GFP-MiR vector.Then the PCI-GFP-MiR-shRNA vector was recombined with the FUW lentiviral vector.The obtained lentiviral vectors containing P450 3A11... |
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| Keywords: | cytochrome P-450 CYP3A knock-down microRNAs lentiviral vector mice |
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