Three-hybrid analysis to study RNA decay S. cerevisiae (i.e, G18 insertions, mutations in nucleases…) Defining degradative activities in Drosophila melanogaster Defining degradative activities in plants RT-PCR product sequencing of mRNA decay intermediates in mammalian cells after decapping and circularization in vitro Tethering Assays Assays of poly(A) tail length (PAT assay, specific oligo-dT anchors…) and polarity of mRNA decay (Invader Assay) In vivo crosslinking-coupled affinity purification of mRNAs for decay analysis (Streptavidin, biotin…) Classification of ribonucleases by computational analyses Analysis of P bodies in Saccharomyces cerevisiae Analysis of stress granules and P bodies in mammalian cells, including neurons PMR1-mediated mRNA decay No-go mRNA in Saccharomyces cerevisiae mRNA decay mediated by decay factor adaptors (SLBP, PUF, Staufen1…) Non-stop decay in Saccharomyces cerevisiae Measuring ARE-mediated mRNA decay in cells Measuring ARE-mediated mRNA decay in animals Methods to study viral-mediated influences on mRNA decay Measuring Saccharomyces cerevisiae mitochondrial mRNA half-life Measuring mRNA decay in Arabidopsis mitochondria Measuring mRNA decay in human mitochondria Measuring mRNA decay in chloroplasts • Nonsense-Mediated mRNA Decay (how do you study NMD, what defines an In Saccharomyces cerevisiae In Drosophila melanogaster In Caenorhabditis elegans In plants In mammals • Analysis of Nuclear RNA Decay Measuring nuclear RNA decay in Saccharomyces cerevisiae (includes pre-mRNA, mRNA, rRNA, TRAMP complex…) Measuring nuclear RNA decay in mammals • tRNA Decay Decay of improperly modified tRNA decay in Sacharomyces cerevisiae • rRNA Decay Decay of nonfunctional rRNA in Sacharomyces cerevisiae • RNAi-Mediated mRNA Decay Defining RNAi targets (e.g., Ago knockdowns, identifying complementary siRNA/miRNA…) Studying miRNAs in unicellular organisms (Chlamydamones reinhardtii) Studying miRNAs in unicellular organisms: RNAi-dependent mRNA turnover in fission yeast: Assays for microRNA-mediated mRNA decay in Caenhorabditis elegans Analyses of microRNA function in zebra fish Targeted deletion of cell-type specific microRNAs in mice for developmental studies
Specific complexes of protein and RNA carry out many essential biological functions, including RNA processing, RNA turnover, RNA folding, as well as the translation of genetic information from mRNA into protein sequences. Messenger RNA (mRNA) decay is now emerging as an important control point and a major contributor to gene expression. Continuing identification of the protein factors and cofactors, and mRNA instability elements responsible for mRNA decay allow researchers to build a comprehensive picture of the highly orchestrated processes involved in mRNA decay and its regulation.
Covers the nonsense-mediated mRNA decay (NMD) or mRNA surveillance pathway Expert researchers introduce the most advanced technologies and techniques to identify mRNA processing, transport, localization and turnover, which are central to the process of gene expression Offers step-by-step lab instructions, including necessary equipment and reagents
Biochemists and researchers in related life science disciplines.
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- © Academic Press 2008
- 15th December 2008
- Academic Press
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University of Rochester, NY, USA
Rutgers University, New Brunswick, NJ, USA