Premium
Optimization of reference gene panels for gene expression analysis in preclinical models of inflammatory skin diseases
Author(s) -
Braun Anne,
Sezin Tanya,
Bezdek Siegfried,
Doxastaki Iosifina,
Zillikens Detlef,
Busch Hauke S.,
Sadik Christian D.
Publication year - 2019
Publication title -
experimental dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.108
H-Index - 96
eISSN - 1600-0625
pISSN - 0906-6705
DOI - 10.1111/exd.13989
Subject(s) - reference genes , gene expression , gene , normalization (sociology) , reverse transcriptase , gene expression profiling , biology , computational biology , genetics , polymerase chain reaction , sociology , anthropology
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.