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Published on: September 14, 2018
Study of MicroRNAs Expression in Food Allergy
Emilio Nuñez-Borque1, Lucía Palacio-Garcia1, Sergio Fernández-Bravo1
1Department of Allergy and Immunology, IIS-Fundación Jiménez Díaz, Universidad Autónoma de Madrid, Madrid, Spain.
Abstract:
Food allergy is a current worldwide problem. Consequently, it is necessary to identify the different molecules that modulate these reactions and that can be used as potential biomarkers. In recent years there has been increasing interest in the field of allergy on microRNAs (miRNAs). These molecules regulate a wide variety of physiological processes and have been proposed as promising candidate biomarkers.Currently, next-generation sequencing (NGS) has allowed to determine the profile of all miRNAs from different samples. In addition, there are several methods to extract RNA and miRNAs, from different sources such as serum, extracellular vesicles (EVs), and/or cell extracts. Following extraction, a retrotranscription step must be carried out before miRNA levels can be quantified by quantitative polymerase chain reaction (qPCR).This chapter aimed to describe the discovery techniques used to determine the differential profile of miRNAs from different types of samples, as well as the diverse methods employed to extract these molecules and quantify specific changes in their levels by qPCR.
Insights
Food allergies are a global concern. Researchers are exploring microRNAs (miRNAs) as potential biomarkers, using next-generation sequencing and qPCR to identify and quantify these molecules for diagnostic purposes.
Area of Science:
- Immunology and Molecular Biology
- Biomarker Discovery
Background:
- Food allergy presents a significant global health challenge.
- MicroRNAs (miRNAs) are increasingly recognized for their role in regulating physiological processes and as potential allergy biomarkers.
- Identifying reliable biomarkers is crucial for understanding and managing food allergies.
Purpose of the Study:
- To outline techniques for discovering differential microRNA (miRNA) profiles in various sample types.
- To describe diverse methods for RNA and miRNA extraction from biological samples.
- To detail the quantification of specific miRNA level changes using quantitative polymerase chain reaction (qPCR).
Main Methods:
- Next-generation sequencing (NGS) for comprehensive miRNA profiling.
- Multiple RNA/miRNA extraction protocols from serum, extracellular vesicles (EVs), and cell extracts.
- Reverse transcription followed by quantitative polymerase chain reaction (qPCR) for miRNA quantification.
Main Results:
- NGS enables the determination of global miRNA profiles across different samples.
- Various extraction methods are available for isolating miRNAs from diverse biological sources.
- qPCR allows for the precise quantification of specific miRNA expression levels.
Conclusions:
- This chapter provides a comprehensive overview of miRNA discovery and quantification techniques relevant to food allergy research.
- The described methods facilitate the identification of potential miRNA biomarkers for food allergy.
- Understanding miRNA modulation is key to advancing diagnostic and therapeutic strategies in allergy.
