The mutation detections of KRAS and BRAF genes are of significant importance to predict the responses to anti-cancer therapy and develop new drugs. In this paper, we developed a multi-step fluorescence resonance energ...The mutation detections of KRAS and BRAF genes are of significant importance to predict the responses to anti-cancer therapy and develop new drugs. In this paper, we developed a multi-step fluorescence resonance energy transfer (FRET) assay for multiplex detection of KRAS and BRAF mutations using cationic conjugated polymers (CCP). The newly established detection system could detect as low as 2% mutant DNAs in DNA admixtures. By triggering the emission intensity change of CCP and the dyes labeled in the DNA, four possible statuses (three mutations and one wildtype) can be differentiated in one extension reaction. The detection efficiency of this new method in clinical molecular diagnosis was validated by determining KRAS and BRAF mutations of 51 formalin-fixed paraffin-embedded (FFPE) ovary tissue samples. Furthermore, the result of the CCP-based multi-step FRET assay can be directly visualized under UV light so that no expensive instruments and technical expertise are needed. Thus, the assay provides a sensitive, reliable, cost-effective and simple method for the detection of disease-related gene mutations.展开更多
A cationic dendritic polyfluorene (PFP) is examined as a siRNA delivery vector. This material was designed to facilitate the nucleic acid binding, encapsulation and efficient cellular uptake. PFP can effectively prote...A cationic dendritic polyfluorene (PFP) is examined as a siRNA delivery vector. This material was designed to facilitate the nucleic acid binding, encapsulation and efficient cellular uptake. PFP can effectively protect siRNA against nuclease degradation, which is necessary for gene carriers. PFP can be used for multidrug resistance gene-targeted siRNA delivery in doxorubicin (Dox)-resistant human breast cancer cells (MCF7) cells. As a siRNA transfection agent, PFP can efficiently achieve the reversal of drug resistance and enhance the drug sensitivity. These new features and capabilities represent a major step toward conjugated polymers that can function for therapeutic application.展开更多
A new strategy is performed to fabricate conjugated polymer microarray with the assistance of protein in this work.The water-soluble cationic conjugated polymer employed in the present work is capable of absorbing lig...A new strategy is performed to fabricate conjugated polymer microarray with the assistance of protein in this work.The water-soluble cationic conjugated polymer employed in the present work is capable of absorbing light at 510 nm,which makes it compatible with a variety of commercial microarray scanners.It is demonstrated that the protein-assisted conjugated polymer microarray exhibits higher fluorescence signal and better stability in comparison with the case without protein.The conjugated polymer microarray can be used for sensitive detection of picric acid(PA).A major advantage of our approach is its simplicity and chemical linking is not required between the conjugated polymer and microarray substrate.Considering the simplicity of the preparation of the conjugated polymer microarray,it is anticipated that novel sensing platforms will be constructed by employing this versatile method.展开更多
基金supported by the National Natural Science Foundation of China (90913014, 21021091)
文摘The mutation detections of KRAS and BRAF genes are of significant importance to predict the responses to anti-cancer therapy and develop new drugs. In this paper, we developed a multi-step fluorescence resonance energy transfer (FRET) assay for multiplex detection of KRAS and BRAF mutations using cationic conjugated polymers (CCP). The newly established detection system could detect as low as 2% mutant DNAs in DNA admixtures. By triggering the emission intensity change of CCP and the dyes labeled in the DNA, four possible statuses (three mutations and one wildtype) can be differentiated in one extension reaction. The detection efficiency of this new method in clinical molecular diagnosis was validated by determining KRAS and BRAF mutations of 51 formalin-fixed paraffin-embedded (FFPE) ovary tissue samples. Furthermore, the result of the CCP-based multi-step FRET assay can be directly visualized under UV light so that no expensive instruments and technical expertise are needed. Thus, the assay provides a sensitive, reliable, cost-effective and simple method for the detection of disease-related gene mutations.
基金supported by the National Natural Science Foundation of China(21203213,21033010,21021091)
文摘A cationic dendritic polyfluorene (PFP) is examined as a siRNA delivery vector. This material was designed to facilitate the nucleic acid binding, encapsulation and efficient cellular uptake. PFP can effectively protect siRNA against nuclease degradation, which is necessary for gene carriers. PFP can be used for multidrug resistance gene-targeted siRNA delivery in doxorubicin (Dox)-resistant human breast cancer cells (MCF7) cells. As a siRNA transfection agent, PFP can efficiently achieve the reversal of drug resistance and enhance the drug sensitivity. These new features and capabilities represent a major step toward conjugated polymers that can function for therapeutic application.
基金supported by the National Natural Science Foundation of China(21203213,21021091)the National Basic Research Program of China(2011CB808400,2013CB932800)
文摘A new strategy is performed to fabricate conjugated polymer microarray with the assistance of protein in this work.The water-soluble cationic conjugated polymer employed in the present work is capable of absorbing light at 510 nm,which makes it compatible with a variety of commercial microarray scanners.It is demonstrated that the protein-assisted conjugated polymer microarray exhibits higher fluorescence signal and better stability in comparison with the case without protein.The conjugated polymer microarray can be used for sensitive detection of picric acid(PA).A major advantage of our approach is its simplicity and chemical linking is not required between the conjugated polymer and microarray substrate.Considering the simplicity of the preparation of the conjugated polymer microarray,it is anticipated that novel sensing platforms will be constructed by employing this versatile method.
