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MicroRNAs

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...

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Related Experiment Video

Updated: Aug 23, 2025

Oral Biofilm Sampling for Microbiome Analysis in Healthy Children

Oral Biofilm Sampling for Microbiome Analysis in Healthy Children

Published on: December 31, 2017

Confounding Factors Impacting microRNA Expression in Human Saliva: Methodological and Biological Considerations.

Rhea Sullivan¹, Austin Montgomery¹, Anna Scipioni^1,2

¹Department of Pediatrics, Penn State Hershey College of Medicine, Hershey, PA 17033, USA.

|October 27, 2022

Summary

Saliva microRNA (miRNA) analysis requires standardized methods. This study identified key experimental factors like RNA stabilizers and collection techniques that significantly impact miRNA expression, crucial for reliable disease biomarker development.

Keywords:

biomarker housekeeping miRNA salivary stabilizer

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Last Updated: Aug 23, 2025

Oral Biofilm Sampling for Microbiome Analysis in Healthy Children

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Collection and Extraction of Saliva DNA for Next Generation Sequencing

Collection and Extraction of Saliva DNA for Next Generation Sequencing

Published on: August 27, 2014

Absolute Quantification of Plasma MicroRNA Levels in Cynomolgus Monkeys, Using Quantitative Real-time Reverse Transcription PCR

Absolute Quantification of Plasma MicroRNA Levels in Cynomolgus Monkeys, Using Quantitative Real-time Reverse Transcription PCR

Published on: February 12, 2018

Area of Science:

Biomarker Discovery
Molecular Diagnostics
Genomics

Background:

Saliva microRNAs (miRNAs) show promise as non-invasive biomarkers for various human diseases.
Reproducibility and generalizability of salivary miRNA studies are hindered by technical variations and inter-individual differences.
A lack of standardized control miRNAs presents a significant challenge in salivary miRNA biomarker research.

Purpose of the Study:

To quantify the variability in miRNA expression in saliva due to different experimental parameters and biological factors.
To identify optimal experimental conditions for salivary miRNA measurement.
To discover reliable reference miRNAs for salivary biomarker studies.

Main Methods:

Analysis of 1225 saliva samples from 922 healthy individuals.
Quantification of miRNA expression variability influenced by aligner selection (Bowtie1 vs. Bowtie2), saliva collection methods (expectorated vs. swabbed), and RNA stabilizer presence/absence.
Assessment of biological factors including sex, age, BMI, exercise, and caloric intake.

Main Results:

Absence of RNA stabilizer caused the most significant variability in miRNA expression.
Saliva collection methods and aligner choice also introduced notable variability.
Biological factors influenced a smaller subset of miRNAs, with coefficients of variation reported for 643 consistently detected miRNAs.

Conclusions:

Standardizing experimental parameters, particularly RNA stabilization and collection methods, is critical for reproducible salivary miRNA biomarker studies.
Several consistently detected salivary miRNAs can serve as effective reference genes.
This research provides a foundation for optimizing salivary miRNA measurement protocols and identifying appropriate control miRNAs.