RNA structure analysis of viruses using SHAPE.

Curr Protoc Microbiol

Department of Microbiology and Immunology, University of California, San Francisco, California.

Published: October 2013

Selective 2' hydroxyl acylation analyzed by primer extension (SHAPE) provides a means to investigate RNA structure with better resolution and higher throughput than has been possible with traditional methods. We present several protocols, which are based on a variety of previously published methods and were adapted and optimized for the analysis of poliovirus RNA in the Andino laboratory. These include methods for nondenaturing RNA extraction, RNA modification and primer extension, and data processing in ShapeFinder.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4826560PMC
http://dx.doi.org/10.1002/9780471729259.mc15h03s30DOI Listing

Publication Analysis

Top Keywords

rna structure
8
primer extension
8
rna
5
structure analysis
4
analysis viruses
4
viruses shape
4
shape selective
4
selective hydroxyl
4
hydroxyl acylation
4
acylation analyzed
4

Similar Publications

Circular RNAs (circRNAs), as a class of noncoding RNA molecules with a circular structure exhibit high stability and spatiotemporal-specific expression, making them ideal cancer biomarkers for liquid biopsy. Herein, a new photoelectrochemical (PEC) biosensor for a highly sensitive circRNA assay in the whole blood of lung cancer patients was designed based on CRISPR/Cas13a-programmed Cu nanoclusters (Cu NCs) and a -scheme covalent organic framework/silver sulfide (T-COF/AgS) composite. This -scheme T-COF/AgS composite accelerates electron transfer and produces an excellent initial photocurrent.

View Article and Find Full Text PDF

The methylation- demethylation dynamics of RNA plays major roles in different biological functions, including stress responses, in plants. mA methylation in RNA is orchestrated by a coordinated function of methyl transferases (writers) and demethylases (Erasers). Genome-wide analysis of genes involved in methylation and demethylation was performed in pigeon pea.

View Article and Find Full Text PDF

DNA Nanotags for Multiplexed Single-Particle Electron Microscopy and Electron Cryotomography.

JACS Au

January 2025

CAS Key Laboratory of Nanosystem and Hierarchical Fabrication, CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology of China, CAS, Beijing 100190, China.

DNA nanostructures present new opportunities as Nanotags for electron microscopy (EM) imaging, leveraging their high programmability, unique shapes, biomolecule conjugation capability, and stability compatible with standard cryogenic sample preparation protocols. This perspective highlights the potential of DNA Nanotags to enable high-throughput multiplexed EM analysis and facilitate particle identification for cryogenic electron tomography (cryo-ET). Meanwhile, applying Nanotags in live-cell environments requires the efficient cellular uptake of intact structures and successful cytosolic migration.

View Article and Find Full Text PDF

Consistent features observed in structural probing data of eukaryotic RNAs.

NAR Genom Bioinform

March 2025

Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwanoha 5-1-5, Kashiwa, Chiba 277-8561, Japan.

Understanding RNA structure is crucial for elucidating its regulatory mechanisms. With the recent commercialization of messenger RNA vaccines, the profound impact of RNA structure on stability and translation efficiency has become increasingly evident, underscoring the importance of understanding RNA structure. Chemical probing of RNA has emerged as a powerful technique for investigating RNA structure in living cells.

View Article and Find Full Text PDF

Directed evolution of peroxidase DNAzymes by a function-based approach.

Biol Methods Protoc

December 2024

School of Biomedical Sciences, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, P.R. China.

Peroxidase DNAzymes are single-stranded, stable G-quadruplexes structures that exhibit catalytic activity with cofactor hemin. This class of DNAzymes offers several advantages over traditional protein and RNA catalysts, including thermal stability, resistance to hydrolysis, and easy of synthesis in the laboratory. However, their use in medicine, biology, and chemistry is limited due to their low catalytic rates.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!