Monitoring p21 mRNA expression in living cell based on molecular beacon fluorescence increasing rate |
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Authors: | Tang HongXing Yang XiaoHai Wang KeMin Tan Weihong Liu Bin He Lifang Wang Wei |
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Institution: | (1) State Key Laboratory of Chemo/Biosensing and Chemometrics, Biomedical Engineering Center, College of Chemistry and Chemical Engineering, and Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha, 410082, China |
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Abstract: | Studying the expression level of mRNA in living cells will offer tremendous opportunities for ad-vancement in cell biology research,disease diagnostics,and drug discovery.In this paper,a molecular beacon(MB)specific for the important tumor suppressor gene p21 has been designed and synthesized.The fluorescence signal was detected in real-time after the MB entered the cytoplasm of nasopharyn-geal carcinoma cells.After injecting the p21MB into nasopharyngeal carcinoma cell and p33-trans-fected nasopharyngeal carcinoma cell,the consistent increase of fluorescent signal intensity was de-tected in both cell lines,and maximum fluorescence intensity achieved in about 15 min.In about 4 min following microinjection,the fluorescence increasing rate was significantly different between these two cell lines,which indicate the different p21 mRNA expression levels.The results obtained in the real-time detection were also validated by RT-PCR.Analysis of the initial fluorescence increasing rate can effi-ciently reduce the side effect of enzyme and improve the accuracy in living cell mRNA detection. |
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Keywords: | molecular beacon p21 mRNA nasopharyngeal carcinoma cell living cell imaging fluorescence increasing rate |
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