MicroRNA Detection and Target Identification: Methods and Protocols

This updated volume reflects new and evolved techniques to study detection, profiling, and manipulation of microRNAs (miRNAs) in plants and animals. It includes introductions to their respective topics, lists of necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Format: Hardback
Length: 266 pages
Publication date: 24 January 2023
Publisher: Springer-Verlag New York Inc.

This updated volume reflects new and evolved techniques to study detection, profiling, and manipulation of microRNAs (miRNAs) in plants and animals. After overviews of how best to detect, identify, and validate microRNAs, the book continues by exploring state-of-the-art protocols for microRNA detection, approaches to profile the expression level of microRNAs, spatial expression analysis, in silico analysis of microRNAs and their targets, as well as protocols for functional analysis of microRNAs and their targets by CRISPR/Cas. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, MicroRNA Detection and Target Identification: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.

MicroRNAs (miRNAs) are small, non-coding RNAs that play crucial roles in regulating gene expression in plants and animals. The study of miRNAs has gained significant attention in recent years due to their important roles in various biological processes, such as development, differentiation, and disease. In this updated volume, we aim to provide a comprehensive and up-to-date overview of the techniques used to detect, profile, and manipulate microRNAs in plants and animals.

Detection of microRNAs:

MicroRNA detection is a critical step in understanding the role of miRNAs in a particular cell or tissue. There are several methods available for microRNA detection, including microarray analysis, reverse transcription-quantitative PCR (RT-qPCR), and next-generation sequencing (NGS). Microarray analysis is a widely used method for profiling the expression of thousands of genes simultaneously, making it an efficient tool for identifying differentially expressed microRNAs. RT-qPCR is a highly sensitive and specific method for quantifying the expression of microRNAs, allowing for accurate measurement of their abundance in different samples. NGS is a powerful technique that enables the sequencing of entire genomes or transcriptomes, providing a comprehensive view of microRNA expression at the whole-genome level.

Profiling of microRNA Expression:

Once microRNAs have been detected, it is important to profile their expression levels in different tissues or cell types. This can be achieved by RT-qPCR or NGS, which allow for the quantification of microRNA abundance in a specific sample. Profiling of microRNA expression can provide valuable insights into the regulatory networks and pathways involved in a particular biological process.

Spatial Expression Analysis:

MicroRNAs are known to play important roles in regulating gene expression at the spatial level. Spatial expression analysis is a technique used to study the distribution of microRNAs in different tissues or cell types. This can be achieved by fluorescent in situ hybridization (FISH), which allows for the visualization of microRNA expression in living cells. FISH can provide information about the localization of microRNAs within cells and tissues, which can be useful in understanding their biological functions.

In Silico Analysis of MicroRNAs and Their Targets:

In silico analysis is a powerful tool for predicting the targets of microRNAs and understanding their biological functions. This can be achieved by using bioinformatics tools, such as miRanda, TargetScan, and PicTar. These tools allow for the identification of potential microRNA targets based on their sequence similarity to known genes. In silico analysis can also help identify microRNA-mRNA interactions, which can provide insights into the regulatory networks involved in a particular biological process.

Functional Analysis of MicroRNAs and Their Targets by CRISPR/Cas:

CRISPR/Cas is a powerful tool for editing the genomes of plants and animals. It has been used to study the function of microRNAs and their targets in a variety of contexts. Functional analysis of microRNAs and their targets by CRISPR/Cas can provide insights into the mechanisms underlying a particular biological process. This can be achieved by knocking out or overexpressing microRNAs or their targets in cells or tissues, and then studying the resulting phenotypes.

Conclusion:

In conclusion, this updated volume reflects new and evolved techniques to study detection, profiling, and manipulation of microRNAs in plants and animals. By exploring state-of-the-art protocols for microRNA detection, approaches to profile the expression level of microRNAs, spatial expression analysis, in silico analysis of microRNAs and their targets, and protocols for functional analysis of microRNAs and their targets by CRISPR/Cas, we aim to provide a comprehensive and up-to-date overview of this vital field. We hope that this book will be a valuable resource for researchers and scientists interested in studying microRNAs and their roles in biological processes.

Weight: 722g
Dimension: 254 x 178 (mm)
ISBN-13: 9781071629819
Edition number: 2nd ed. 2023