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.
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http://dx.doi.org/10.1002/9780471729259.mc15h03s30 | DOI Listing |
ACS Sens
January 2025
College of Public Health, Zhengzhou University, Zhengzhou 450001, China.
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 PDFFront Plant Sci
January 2025
National Institute of Plant Biotechnology, Indian Council of Agricultural Research (ICAR), New Delhi, India.
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 PDFJACS 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 PDFNAR 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 PDFBiol 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.
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