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shRNA knock-down efficiency test in vitro and transgenic founder mouse production.

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posted on 2013-02-20, 09:34 authored by Yong Wang, Hai-Hong Hu, Hao Pang, Xiao-Yang Zhou, Lu-Shan Yu, Lu-Lu Wang, Cang'e Liu, Ke-Nan Guo, Cong Zhao, Qin Liu, Ben-Hua Zeng, Huan Tang, Hai-Tao Shang, Su Zeng, Hong Wei

A: CYP3A expression was detected by RT-PCR in hepatic cells expressing the two designed shRNAs (shRNA1 and shRNA2), shRNA targeting luciferase gene (negative control) and untreated hepatic cells (blank control) respectively. 1: DNA markers; 2: untreated hepatic cells (blank control); 3: hepatic cells infected with an unrelated shRNA targeting luciferase gene (negative control); 4: hepatic cells infected with shRNA1; 5: hepatic cells infected with shRNA2. B: The CYP3A expression level in hepatic cells expressing shRNA1 or shRNA2 was significantly lower than that of negative or blank control, and hepatic cells expressing shRNA1 exhibited the lowest CYP3A expression level. C: Transgenic mice were generated with the lentiviral vector expressing shRNA1, and four founder mice were detected to be transgenic by PCR using genomic DNA as templates. D. The four transgenic founder mice displayed fluorescence of different intensities. *: indicates statistic significance.