کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
11016384 1777900 2019 21 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A novel dual energy transfer probe for intracellular mRNA detection with high robustness and specificity
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
A novel dual energy transfer probe for intracellular mRNA detection with high robustness and specificity
چکیده انگلیسی
Real-time detection and visualization of the content and distribution of mRNA in living cells are important in molecular biology and can provide information about cell heterogeneity for clinical studies. Traditional mRNA detection methods require cell lysis to extract mRNA, which is complicated and cannot guarantee cell viability or the physiological function of the target nucleic acids. We propose a versatile fluorescent probe based on oligonucleotide displacement and dual fluorescence resonance energy transfer (FRET) for intracellular mRNA measurements. Two types of FRET probes, the Cy3 probe and Cy5 probe, were synthesized using gold nanoparticles (AuNPs) as the quencher and a double-stranded DNA as the bridge between fluorophores and AuNPs. Both FRET probes were able to label target mRNA. Energy transfer between Cy3 and Cy5 only occurs when both FRET probes label target mRNA after oligonucleotide displacement. Based on intracellular mRNA detection and selectivity and cytotoxicity studies, this method was shown to be able to determine the localization and expression level of mRNA in living cells with high robustness and specificity. Further investigation of the influence of anti-tumor drugs on the transcription level of the survivin gene illustrated the potential application of this method in drug screening and disease studies.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Sensors and Actuators B: Chemical - Volume 279, 15 January 2019, Pages 342-350
نویسندگان
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