TY - JOUR
T1 - Optimization of reference gene panels for gene expression analysis in preclinical models of inflammatory skin diseases
AU - Braun, Anne
AU - Sezin, Tanya
AU - Bezdek, Siegfried
AU - Doxastaki, Iosifina
AU - Zillikens, Detlef
AU - Busch, Hauke S
AU - Sadik, Christian D
N1 - © 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
PY - 2019/8/1
Y1 - 2019/8/1
N2 - Reverse transcriptase qPCR is the most common method to determine and compare mRNA expression levels and relies on normalization using reference genes. The expression levels of the latter, however, are themselves often variable between experimental conditions, thus compromising the results. Using the geNorm algorithm, we have examined seven genes with respect to their suitability as reference genes for gene analysis in mouse models of skin inflammation, using the antibody transfer model of epidermolysis bullosa acquisita and in the Aldara™ -induced psoriasiform dermatitis. Our results indicate that the combination of at least 2-3 reference genes is required for stable normalization. Notably, the expression of reference genes changed when comparing lesional skin of both models or when comparing lesional to non-lesional skin within one model. This highlights the need for precise selection of reference genes dependent on the specific experimental setup.
AB - Reverse transcriptase qPCR is the most common method to determine and compare mRNA expression levels and relies on normalization using reference genes. The expression levels of the latter, however, are themselves often variable between experimental conditions, thus compromising the results. Using the geNorm algorithm, we have examined seven genes with respect to their suitability as reference genes for gene analysis in mouse models of skin inflammation, using the antibody transfer model of epidermolysis bullosa acquisita and in the Aldara™ -induced psoriasiform dermatitis. Our results indicate that the combination of at least 2-3 reference genes is required for stable normalization. Notably, the expression of reference genes changed when comparing lesional skin of both models or when comparing lesional to non-lesional skin within one model. This highlights the need for precise selection of reference genes dependent on the specific experimental setup.
UR - http://www.scopus.com/inward/record.url?scp=85068621864&partnerID=8YFLogxK
UR - http://www.mendeley.com/research/optimization-reference-gene-panels-gene-expression-analysis-preclinical-models-inflammatory-skin-dis
U2 - 10.1111/exd.13989
DO - 10.1111/exd.13989
M3 - Letters
C2 - 31177560
SN - 0906-6705
VL - 28
SP - 985
EP - 988
JO - Experimental Dermatology
JF - Experimental Dermatology
IS - 8
ER -