Enzymes in RNA Science and Biotechnology

Enzymes in RNA Science and Biotechnology,Volume 691,covers new advances in reverse transcriptase, structured RNAs, RNA G-quadruplexes, tRNA structure-seq, and CAP-STRUCTURE-seq, providing valuable insights into RNA research and biotechnology.

Format: Hardback
Length: 412 pages
Publication date: 01 November 2023
Publisher: Elsevier Science Publishing Co Inc

Enzymes in RNA Science and Biotechnology,Volume 691 in the Methods in Enzymology series, presents a comprehensive overview of the latest advancements in the field, encompassing chapters on Reverse transcriptase Part I (discovery, preparation, general utilization, MarathonRT for routine RT-PCR and for cDNA synthesis on challenging RNA templates), Structured RNAs, repeat RNAs, and more. The book also delves into Engineering TNA Polymerases Through Iterative Cycles of Directed Evolution, Reverse transcriptase Part II (RNA structure mapping and determination), RNA G-quadruplex (rG4) structure detection using RTS and SHALiPE assays, tRNA Structure-seq in vivo and in droplets, Capture the in vivo intact RNA structurome by CAP-STRUCTURE-seq, and other groundbreaking techniques.

This comprehensive volume serves as a valuable resource for researchers and practitioners in RNA Science and Biotechnology, providing a detailed account of the latest methodologies and techniques employed in this rapidly evolving field. The chapters authored by renowned experts in the field offer insightful perspectives and practical insights, making it an essential tool for anyone seeking to stay updated with the latest developments in RNA research.

The book's organization is well-structured, with each chapter presenting a clear and concise introduction to the topic, followed by detailed explanations, examples, and references. The use of illustrative figures, tables, and diagrams further enhances the understanding of the subject matter, making it accessible to a wide range of readers, including students, researchers, and professionals.

One of the key highlights of this book is its coverage of Reverse transcriptase Part I. The chapter provides a comprehensive overview of the discovery, preparation, general utilization, and specific applications of reverse transcriptase in RNA research. It discusses the various types of reverse transcriptases available, their characteristics, and their applications in RNA isolation, cDNA synthesis, and RNA structure mapping. The authors also provide detailed protocols for the preparation of reverse transcriptase and its optimization for different RNA templates.

Another notable chapter is Engineering TNA Polymerases Through Iterative Cycles of Directed Evolution. This chapter explores the use of directed evolution to engineer TNA polymerases, which are essential enzymes involved in RNA synthesis. The authors describe the iterative cycles of directed evolution, which involve mutagenesis, selection, and screening, to improve the catalytic efficiency and specificity of TNA polymerases. The chapter also provides examples of engineered TNA polymerases with enhanced properties, such as increased fidelity and tolerance to nucleotide mismatches.

The book also delves into RNA G-quadruplex (rG4) structure detection using RTS and SHALiPE assays. The rG4 structure is a unique RNA secondary structure that plays important roles in gene regulation and RNA processing. The authors discuss the methods used to detect rG4 structures, including RTS and SHALiPE assays, and their applications in RNA research. The chapter provides detailed protocols for the preparation of RNA samples, the execution of the assays, and the interpretation of the results.

In addition to these chapters, the book also includes a chapter on tRNA Structure-seq in vivo and in droplets. tRNA is a crucial component of the ribosome, responsible for translating mRNA into amino acids. The authors discuss the methods used to sequence tRNA in vivo and in droplets, including single-molecule sequencing and droplet-based sequencing technologies. The chapter provides detailed protocols for the preparation of RNA samples, the execution of the sequencing experiments, and the analysis of the data.

Another important chapter is Capture the in vivo intact RNA structurome by CAP-STRUCTURE-seq. CAP-STRUCTURE-seq is a novel method that allows the simultaneous mapping of RNA structures and the identification of RNA-protein interactions in living cells. The authors discuss the principles of CAP-STRUCTURE-seq, the experimental setup, and the analysis of the data. The chapter provides detailed protocols for the preparation of RNA samples, the execution of the sequencing experiments, and the interpretation of the results.

Overall, Enzymes in RNA Science and Biotechnology,Volume 691 in the Methods in Enzymology series, is a comprehensive and authoritative resource for researchers and practitioners in RNA Science and Biotechnology. The book presents a wide range of cutting-edge techniques and methodologies, covering topics such as reverse transcriptase, structured RNAs, repeat RNAs, engineering TNA polymerases, RNA G-quadruplex structure detection, tRNA Structure-seq, and Capture the in vivo intact RNA structurome. The chapters authored by renowned experts in the field offer insightful perspectives and practical insights, making it an essential tool for anyone seeking to advance their knowledge in RNA research.

Dimension: 229 x 151 (mm)
ISBN-13: 9780443157707