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Expressed repetitive elements are broadly applicable reference targets for normalization of reverse transcription-qPCR data in mice

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Abstract
Reverse transcription quantitative PCR (RT-qPCR) is the gold standard method for gene expression analysis on mRNA level. To remove experimental variation, expression levels of the gene of interest are typically normalized to the expression level of stably expressed endogenous reference genes. Identifying suitable reference genes and determining the optimal number of reference genes should precede each quantification study. Popular reference genes are not necessarily stably expressed in the examined conditions, possibly leading to inaccurate results. Stably and universally expressed repetitive elements (ERE) have previously been shown to be an excellent alternative for normalization using classic reference genes in human and zebrafish samples. Here, we confirm that in mouse tissues, EREs are broadly applicable reference targets for RT-qPCR normalization, provided that the RNA samples undergo a thorough DNase treatment. We identified Orr1a0, Rltr2aiap, and Rltr13a3 as the most stably expressed mouse EREs across six different experimental conditions. Therefore, we propose this set of ERE reference targets as good candidates for normalization of RT-qPCR data in a plethora of conditions. The identification of widely applicable stable mouse RT-qPCR reference targets for normalization has great potential to facilitate future murine gene expression studies and improve the validity of RT-qPCR data.
Keywords
REAL-TIME PCR, GENE-EXPRESSION, MARFAN-SYNDROME, IN-VIVO, DIFFERENTIATION, MODEL

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Chicago
Renard, Marjolijn, Suzanne Vanhauwaert, Marine Vanhomwegen, Ali Rihani, Niels Vandamme, Steven Goossens, Geert Berx, et al. 2018. “Expressed Repetitive Elements Are Broadly Applicable Reference Targets for Normalization of Reverse transcription-qPCR Data in Mice.” Scientific Reports 8.
APA
Renard, M., Vanhauwaert, S., Vanhomwegen, M., Rihani, A., Vandamme, N., Goossens, S., Berx, G., et al. (2018). Expressed repetitive elements are broadly applicable reference targets for normalization of reverse transcription-qPCR data in mice. SCIENTIFIC REPORTS, 8.
Vancouver
1.
Renard M, Vanhauwaert S, Vanhomwegen M, Rihani A, Vandamme N, Goossens S, et al. Expressed repetitive elements are broadly applicable reference targets for normalization of reverse transcription-qPCR data in mice. SCIENTIFIC REPORTS. 2018;8.
MLA
Renard, Marjolijn, Suzanne Vanhauwaert, Marine Vanhomwegen, et al. “Expressed Repetitive Elements Are Broadly Applicable Reference Targets for Normalization of Reverse transcription-qPCR Data in Mice.” SCIENTIFIC REPORTS 8 (2018): n. pag. Print.
@article{8581135,
  abstract     = {Reverse transcription quantitative PCR (RT-qPCR) is the gold standard method for gene expression analysis on mRNA level. To remove experimental variation, expression levels of the gene of interest are typically normalized to the expression level of stably expressed endogenous reference genes. Identifying suitable reference genes and determining the optimal number of reference genes should precede each quantification study. Popular reference genes are not necessarily stably expressed in the examined conditions, possibly leading to inaccurate results. Stably and universally expressed repetitive elements (ERE) have previously been shown to be an excellent alternative for normalization using classic reference genes in human and zebrafish samples. Here, we confirm that in mouse tissues, EREs are broadly applicable reference targets for RT-qPCR normalization, provided that the RNA samples undergo a thorough DNase treatment. We identified Orr1a0, Rltr2aiap, and Rltr13a3 as the most stably expressed mouse EREs across six different experimental conditions. Therefore, we propose this set of ERE reference targets as good candidates for normalization of RT-qPCR data in a plethora of conditions. The identification of widely applicable stable mouse RT-qPCR reference targets for normalization has great potential to facilitate future murine gene expression studies and improve the validity of RT-qPCR data.},
  articleno    = {7642},
  author       = {Renard, Marjolijn and Vanhauwaert, Suzanne and Vanhomwegen, Marine and Rihani, Ali and Vandamme, Niels and Goossens, Steven and Berx, Geert and Van Vlierberghe, Pieter and Haigh, Jody J and Decaesteker, Bieke and Van Laere, Jolien and Lambertz, Irina and Speleman, Franki and Vandesompele, Jo and Willaert, Andy},
  issn         = {2045-2322},
  journal      = {SCIENTIFIC REPORTS},
  language     = {eng},
  pages        = {9},
  title        = {Expressed repetitive elements are broadly applicable reference targets for normalization of reverse transcription-qPCR data in mice},
  url          = {http://dx.doi.org/10.1038/s41598-018-25389-6},
  volume       = {8},
  year         = {2018},
}

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