AU Content in the MicroRNA Sequence Influences its Stability After Heat Treatment

Agnès Garcias López, Louise Brogaard, Peter M. H. Heegaard, Susanna Cirera, Kerstin Skovgaard*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

BACKGROUND: MicroRNAs (miRNAs) are short non-coding RNA molecules which regulate gene expression post-transcriptionally and are involved in a multitude of cellular processes. MiRNAs are known to be very stable compared to messenger RNAs (mRNAs), making them excellent candidates as biomarkers for disease. Recently, studies have suggested that miRNA stability in formalin fixed samples might dependend on their nucleotide composition.

OBJECTIVE: To explore the stability of a panel of miRNAs isolated from porcine blood and lung tissue after heat and enzyme treatment.

METHOD: Porcine RNA isolated from lung tissue and blood leukocytes were used for this study. RNA samples were exposed to heat treatment and RNAse A digestion. The levels of selected miRNAs were measured by means of qPCR before and after heat and enzyme treatment.

RESULTS: Fourteen miRNAs were successfully analysed, and they were found to degrade at differently after exposure to heat or RNAse A. MiRNAs with <60% of adenine (A) and uracil (U)in their sequence were found to be more stable.

CONCLUSION: This is the first study showing that different miRNAs isolated from lung tissue display unequal stability after heat treatment, probably based on their nucleotide composition, highlighting the importance of considering the miRNA sequence when investigating their value as biomarkers.

Original languageEnglish
JournalMicroRNA
Volume8
Issue number3
Pages (from-to)216-222
Number of pages7
ISSN2211-5366
DOIs
Publication statusPublished - 2019

Keywords

  • % AU
  • Heat treatment
  • miRNA
  • RNAse A digestion
  • Stability
  • Saliva

Cite this

@article{45b86290e25249e394d0e03579161b62,
title = "AU Content in the MicroRNA Sequence Influences its Stability After Heat Treatment",
abstract = "BACKGROUND: MicroRNAs (miRNAs) are short non-coding RNA molecules which regulate gene expression post-transcriptionally and are involved in a multitude of cellular processes. MiRNAs are known to be very stable compared to messenger RNAs (mRNAs), making them excellent candidates as biomarkers for disease. Recently, studies have suggested that miRNA stability in formalin fixed samples might dependend on their nucleotide composition.OBJECTIVE: To explore the stability of a panel of miRNAs isolated from porcine blood and lung tissue after heat and enzyme treatment.METHOD: Porcine RNA isolated from lung tissue and blood leukocytes were used for this study. RNA samples were exposed to heat treatment and RNAse A digestion. The levels of selected miRNAs were measured by means of qPCR before and after heat and enzyme treatment.RESULTS: Fourteen miRNAs were successfully analysed, and they were found to degrade at differently after exposure to heat or RNAse A. MiRNAs with <60{\%} of adenine (A) and uracil (U)in their sequence were found to be more stable.CONCLUSION: This is the first study showing that different miRNAs isolated from lung tissue display unequal stability after heat treatment, probably based on their nucleotide composition, highlighting the importance of considering the miRNA sequence when investigating their value as biomarkers.",
keywords = "{\%} AU, Heat treatment, miRNA, RNAse A digestion, Stability, Saliva",
author = "L{\'o}pez, {Agn{\`e}s Garcias} and Louise Brogaard and Heegaard, {Peter M. H.} and Susanna Cirera and Kerstin Skovgaard",
note = "Copyright{\circledC} Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.",
year = "2019",
doi = "10.2174/2211536608666190131102252",
language = "English",
volume = "8",
pages = "216--222",
journal = "MicroRNA",
issn = "2211-5366",
publisher = "Bentham Science Publishers Ltd.",
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}

AU Content in the MicroRNA Sequence Influences its Stability After Heat Treatment. / López, Agnès Garcias; Brogaard, Louise; Heegaard, Peter M. H.; Cirera, Susanna; Skovgaard, Kerstin.

In: MicroRNA, Vol. 8, No. 3, 2019, p. 216-222.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - AU Content in the MicroRNA Sequence Influences its Stability After Heat Treatment

AU - López, Agnès Garcias

AU - Brogaard, Louise

AU - Heegaard, Peter M. H.

AU - Cirera, Susanna

AU - Skovgaard, Kerstin

N1 - Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

PY - 2019

Y1 - 2019

N2 - BACKGROUND: MicroRNAs (miRNAs) are short non-coding RNA molecules which regulate gene expression post-transcriptionally and are involved in a multitude of cellular processes. MiRNAs are known to be very stable compared to messenger RNAs (mRNAs), making them excellent candidates as biomarkers for disease. Recently, studies have suggested that miRNA stability in formalin fixed samples might dependend on their nucleotide composition.OBJECTIVE: To explore the stability of a panel of miRNAs isolated from porcine blood and lung tissue after heat and enzyme treatment.METHOD: Porcine RNA isolated from lung tissue and blood leukocytes were used for this study. RNA samples were exposed to heat treatment and RNAse A digestion. The levels of selected miRNAs were measured by means of qPCR before and after heat and enzyme treatment.RESULTS: Fourteen miRNAs were successfully analysed, and they were found to degrade at differently after exposure to heat or RNAse A. MiRNAs with <60% of adenine (A) and uracil (U)in their sequence were found to be more stable.CONCLUSION: This is the first study showing that different miRNAs isolated from lung tissue display unequal stability after heat treatment, probably based on their nucleotide composition, highlighting the importance of considering the miRNA sequence when investigating their value as biomarkers.

AB - BACKGROUND: MicroRNAs (miRNAs) are short non-coding RNA molecules which regulate gene expression post-transcriptionally and are involved in a multitude of cellular processes. MiRNAs are known to be very stable compared to messenger RNAs (mRNAs), making them excellent candidates as biomarkers for disease. Recently, studies have suggested that miRNA stability in formalin fixed samples might dependend on their nucleotide composition.OBJECTIVE: To explore the stability of a panel of miRNAs isolated from porcine blood and lung tissue after heat and enzyme treatment.METHOD: Porcine RNA isolated from lung tissue and blood leukocytes were used for this study. RNA samples were exposed to heat treatment and RNAse A digestion. The levels of selected miRNAs were measured by means of qPCR before and after heat and enzyme treatment.RESULTS: Fourteen miRNAs were successfully analysed, and they were found to degrade at differently after exposure to heat or RNAse A. MiRNAs with <60% of adenine (A) and uracil (U)in their sequence were found to be more stable.CONCLUSION: This is the first study showing that different miRNAs isolated from lung tissue display unequal stability after heat treatment, probably based on their nucleotide composition, highlighting the importance of considering the miRNA sequence when investigating their value as biomarkers.

KW - % AU

KW - Heat treatment

KW - miRNA

KW - RNAse A digestion

KW - Stability

KW - Saliva

U2 - 10.2174/2211536608666190131102252

DO - 10.2174/2211536608666190131102252

M3 - Journal article

VL - 8

SP - 216

EP - 222

JO - MicroRNA

JF - MicroRNA

SN - 2211-5366

IS - 3

ER -