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The dual luciferase reporter system and RT-qPCR strategies for screening of MicroRNA-21 small-molecule inhibitors.

Biotechnol. Appl. Biochem.. 2019; 
HeiYuan-Yuan,GuoYuan-Xu,JiangCong-Shan,WangSi,LuShe-Min,ZhangSa
Products/Services Used Details Operation
DNA Sequencing Plasmid construction Single-strand DNA oligonucleotides of the sense and antisense linker DNA fragment were synthesized (Genscript Company, Suzhou, China) and dissolved in pure water at 10 µM.... Success- ful insertion of the designed linker DNA into backbone plasmids was validated by enzyme digestion and sequencing (Genscript, Suzhou, China). Get A Quote

摘要

The therapeutic potential of microRNA-21 (miR-21) small-molecule inhibitors has been of particular interest to medicinal chemists. Moreover, the development of more facile screening methods is lacking. In the present study, two potential screening strategies for miR-21 small-molecule inhibitor including the stem-loop reverse transcription-quantitative PCR and dual luciferase reporter assay system were demonstrated and discussed in detail. A pmirGLO-miR21cswt plasmid and its two different mutants were constructed for dual luciferase reporter assay system. In addition, the sensitivity and specificity of these two methods were validated. Our results demonstrated that both strategies are decent choices for th... More

关键词

RT-qPCR,dual luciferase assay,microRNA-21,plasmid reconstruction,small-molecule inhibitor scree