Identification of suitable reference genes in the mouse placenta

María Emilia Solano; Kristin Thiele; Mirka Katharina Kowal; Petra Clara Arck

doi:10.1016/j.placenta.2015.12.017

Identification of suitable reference genes in the mouse placenta

Standard

Identification of suitable reference genes in the mouse placenta. / Solano, María Emilia; Thiele, Kristin; Kowal, Mirka Katharina; Arck, Petra Clara.

In: PLACENTA, Vol. 39, 03.2016, p. 7-15.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Solano, ME, Thiele, K, Kowal, MK & Arck, PC 2016, 'Identification of suitable reference genes in the mouse placenta', PLACENTA, vol. 39, pp. 7-15. https://doi.org/10.1016/j.placenta.2015.12.017

APA

Solano, M. E., Thiele, K., Kowal, M. K., & Arck, P. C. (2016). Identification of suitable reference genes in the mouse placenta. PLACENTA, 39, 7-15. https://doi.org/10.1016/j.placenta.2015.12.017

Vancouver

Solano ME, Thiele K, Kowal MK, Arck PC. Identification of suitable reference genes in the mouse placenta. PLACENTA. 2016 Mar;39:7-15. https://doi.org/10.1016/j.placenta.2015.12.017

Bibtex

@article{6af3985ccb904f7d97323d01d8cd415c,

title = "Identification of suitable reference genes in the mouse placenta",

abstract = "INTRODUCTION: Quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) is a reliable tool to analyse gene expression profiles. The expression of housekeeping genes generally serves as a reference for mRNA amount, assuming that it remains stable under pathophysiological and experimental conditions. To date, an empirical validation of reference genes suitable for RT-qPCR-based studies in the mouse placenta is missing.METHODS: We used NormFinder and BestKeeper statistical software to analyse the expression stability of candidate housekeeping genes quantified by RT-qPCR in mouse placentas.RESULTS: Fifteen of 32 potential candidate housekeeping genes analysed on gestation day (gd) 16.5 in mouse placentas exhibited an optimal cycle threshold (Ct). Among them B2m, Polr2a, Ubc, and Ywhaz genes showed the highest expression stability in placentas from control, but also experimentally-challenged mice. These genes as well as the currently widely used housekeeping genes Hprt1, Actb, and Gapdh were selected for further quality assessments. We quantified the Ct values of these selected genes in placental samples obtained from wild-type or genetically engineered dams at different gds, or upon selected experimental interventions known to affect placental phenotype. Among all housekeeping genes analysed, Polr2a was the most stably expressed and its expression stability excelled in combination with Ubc.DISCUSSION: Polr2a, especially in combination with Ubc, can be proposed as highly suitable endogenous reference for gene expression analysis in mouse-derived placental tissue. Moreover, the validation of both genes as a stable reference gene in human placenta-derived tissue strengthens the translational relevance of RT-qPCR findings using mouse placenta.",

author = "Solano, {Mar{\'i}a Emilia} and Kristin Thiele and Kowal, {Mirka Katharina} and Arck, {Petra Clara}",

year = "2016",

month = mar,

doi = "10.1016/j.placenta.2015.12.017",

language = "English",

volume = "39",

pages = "7--15",

journal = "PLACENTA",

issn = "0143-4004",

publisher = "W.B. Saunders Ltd",

}

RIS

TY - JOUR

T1 - Identification of suitable reference genes in the mouse placenta

AU - Solano, María Emilia

AU - Thiele, Kristin

AU - Kowal, Mirka Katharina

AU - Arck, Petra Clara

PY - 2016/3

Y1 - 2016/3

N2 - INTRODUCTION: Quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) is a reliable tool to analyse gene expression profiles. The expression of housekeeping genes generally serves as a reference for mRNA amount, assuming that it remains stable under pathophysiological and experimental conditions. To date, an empirical validation of reference genes suitable for RT-qPCR-based studies in the mouse placenta is missing.METHODS: We used NormFinder and BestKeeper statistical software to analyse the expression stability of candidate housekeeping genes quantified by RT-qPCR in mouse placentas.RESULTS: Fifteen of 32 potential candidate housekeeping genes analysed on gestation day (gd) 16.5 in mouse placentas exhibited an optimal cycle threshold (Ct). Among them B2m, Polr2a, Ubc, and Ywhaz genes showed the highest expression stability in placentas from control, but also experimentally-challenged mice. These genes as well as the currently widely used housekeeping genes Hprt1, Actb, and Gapdh were selected for further quality assessments. We quantified the Ct values of these selected genes in placental samples obtained from wild-type or genetically engineered dams at different gds, or upon selected experimental interventions known to affect placental phenotype. Among all housekeeping genes analysed, Polr2a was the most stably expressed and its expression stability excelled in combination with Ubc.DISCUSSION: Polr2a, especially in combination with Ubc, can be proposed as highly suitable endogenous reference for gene expression analysis in mouse-derived placental tissue. Moreover, the validation of both genes as a stable reference gene in human placenta-derived tissue strengthens the translational relevance of RT-qPCR findings using mouse placenta.

AB - INTRODUCTION: Quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) is a reliable tool to analyse gene expression profiles. The expression of housekeeping genes generally serves as a reference for mRNA amount, assuming that it remains stable under pathophysiological and experimental conditions. To date, an empirical validation of reference genes suitable for RT-qPCR-based studies in the mouse placenta is missing.METHODS: We used NormFinder and BestKeeper statistical software to analyse the expression stability of candidate housekeeping genes quantified by RT-qPCR in mouse placentas.RESULTS: Fifteen of 32 potential candidate housekeeping genes analysed on gestation day (gd) 16.5 in mouse placentas exhibited an optimal cycle threshold (Ct). Among them B2m, Polr2a, Ubc, and Ywhaz genes showed the highest expression stability in placentas from control, but also experimentally-challenged mice. These genes as well as the currently widely used housekeeping genes Hprt1, Actb, and Gapdh were selected for further quality assessments. We quantified the Ct values of these selected genes in placental samples obtained from wild-type or genetically engineered dams at different gds, or upon selected experimental interventions known to affect placental phenotype. Among all housekeeping genes analysed, Polr2a was the most stably expressed and its expression stability excelled in combination with Ubc.DISCUSSION: Polr2a, especially in combination with Ubc, can be proposed as highly suitable endogenous reference for gene expression analysis in mouse-derived placental tissue. Moreover, the validation of both genes as a stable reference gene in human placenta-derived tissue strengthens the translational relevance of RT-qPCR findings using mouse placenta.

U2 - 10.1016/j.placenta.2015.12.017

DO - 10.1016/j.placenta.2015.12.017

M3 - SCORING: Journal article

C2 - 26992668

VL - 39

SP - 7

EP - 15

JO - PLACENTA

JF - PLACENTA

SN - 0143-4004

ER -