Titlebook: Imaging Gene Expression; Methods and Protocol Yaron Shav-Tal Book 2019Latest edition Springer Science+Business Media, LLC, part of Springer

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Live-Cell Imaging of Long Noncoding RNAs Using Molecular Beaconsd on molecular beacons (MBs), which are a class of fluorogenic oligonucleotide-based probes with the capacity to convert RNA target hybridization into a measurable fluorescence signal. In this chapter, we describe the detailed protocol of using MBs to illuminate lncRNA transcripts at the single-molecule level in living cells.

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Visualization of Single mRNAs in Live Neuronse neurons. Then, GFP-labeled RNAs in live neurons can be detected by epifluorescence microscopy, and their moving pathways can be analyzed using single-particle tracking software. For these processes, we introduce protocols for neuron culture, transfection, imaging, and particle tracking methods.

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Quantification of mRNA Turnover in Living Cells: A Pipeline for TREAT Data Analysisndividual mRNA particles. It colocalizes tracks and applies the colocalization information to quantify the number of intact transcripts and degradation intermediates. Based on the analysis of control data, the workflow further determines detection efficiencies and uses them to correct RNA particle numbers.

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Detection of mRNAs Anchored to the Nuclear Envelope During Export Inhibition in Living Cells examine their intracellular distribution and dynamics both in the nucleoplasm and at the nuclear periphery. We use this method to identify and count the number of static mRNPs anchored to the nuclear envelope under different conditions of mRNA export inhibition.

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Imaging Single mRNA Molecules in Mammalian Cells Using an Optimized MS2-MCP Systemogical question triggered the development of the MS2-MCP (MS2-Capsid Protein) system to tag mRNAs and image their life cycle using widefield fluorescence microscopy. The last two decades have evolved toward improving the qualitative and quantitative characteristics of the MS2-MCP system. Here, we pr

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