Interferential RNA retrovirus and its preparation and use
A technology for retroviruses and uses, which is applied in the field of interfering RNA retroviruses and their preparation, can solve problems such as losses, and achieve effects of strong specificity, avoiding sequelae, and high technical content
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[0024] 1. Construction of retroviral vectors expressing interfering RNA
[0025] The DNA sequence of the mouse U6 promoter is loaded into a retroviral vector, and the interfering RNA of BCR / ABL (chronic myeloid leukemia) or CCR5 (HIV receptor) is installed downstream of the U6 promoter, which constitutes Retroviral vectors for expressing interfering RNA, such as figure 1 shown. The vector also contains the gene sequence for expressing green protein. Therefore, it can be used as an index to detect the efficiency of cell transfection or virus infection.
[0026] 2. Detect the interference activity of interfering RNA on the target gene
[0027] Using the method of transient transfection, the plasmid vector expressing OCILRP2 and the retroviral vector expressing interfering RNA were co-transfected into 293F cell line such as figure 2 As shown, after 48 hours, the cells were collected, and the expression of the target gene with red fluorescence was detected by flow cytometry a...
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