A method of screening an HIV-1 activity determinative factor based on high-throughput RNAi
A technology of HIV-1, determinant, applied in the field of molecular biology
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[0032] Example 1 Detection of HIV-1 LTR Regulated Transcriptional Activity Participated by Kinases
[0033] The HIV-1 LTR-hRluc / HSV TK-hLuc luciferase dual reporter plasmid was stably expressed in HEK293T by using shRNA library high-throughput RNAi screening technology for 474 kinases; Take the fluorescence intensity of hLuc (such as figure 1 shown in A);
[0034] Two positive controls, NFκB and AKT1, were used for screening as shRNA or specific inhibitors (eg figure 1 Shown in B); inhibitors of NFκB or AKT1 kinase activity inhibited the activity of HIV-1 LTR by more than 30%, indicating that the screening system was reliable; in addition, the initial screening results showed 155 positive samples against HIV-1 LTR inhibited by 136 genes activity over 20% (eg figure 1 Shown in C), then above-mentioned 155 samples enter the second screening system, obtained 15 positive genes, the results are as shown in table 1;
[0035] Table 1. qPCR. Primers
[0036]
[0037] The resul...
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