FLAG tag peptide and the use of lentiviral vectors in mice foxp3 collaborative filtering effective target gene RNA interference.docVIP

 PAGE \* MERGEFORMAT 21 FLAG tag peptide and the use of lentiviral vectors in mice foxp3 collaborative filtering effective target gene RNA interference Authors: Yan-Xia, Wen-Jie Han, Peng soft-jun, Zhang Hang, Hui-Qiang Huang Wen-Qi Jiang 【Abstract】 【Objective】 screening and efficient foxp3 RNA interference target for blocking the CD4 CD25 Treg cells in the immune suppression. 【Methods】 oligoengine software design foxp3 gene shRNA sequence, chemical synthesis synthesis of four shRNA sequences to construct the target sequence containing foxp3 lentiviral vector; construct a FLAG fusion protein expression and a tool for gene-cell 293T cells for screening RNA interference with a valid target; 【Results】 The successful packaging of the four foxp3 gene target sequence corresponding slow virus-like particles; expression of fusion protein was successfully constructed and purpose of tools for gene 293T cells; Tools cells screened out two separate RNAi efficiency was 91.3% and 95.6% of the valid target. 【Conclusion】 using the FLAG fusion proteins and lentiviral vectors can filter out the best target for RNAi gene Foxp3. Keywords: foxp3; FLAG protein; lentiviral vector, CD4 CD25 Treg, RNA interference Abstract: 【Objective】 To screen the high effective RNA intervening targets of foxp3 gene to block the immunosuppression of CD4 CD25 Tregs. 【Method】 Four shRNA sequences were designed by oligoengine software and synthesized by chemical method. Lentiviral vector that consists target sequences of foxp3 was constructed. The 293T tool cell line that express FLAG fusion protein and target gene was constructed to screen the effective targets for RNA interference. 【Result】 Lentiviral vector that contains foxp3 cDNA sequences was constructed successfully. The 293T tool cells that express FLAG fusion protein and foxp3 gene fragments were constructed successfully. Two effective targets for RNAi were screened in 293T tool cells, the blocking efficiency were 9