Evidence based selection of housekeeping genes.
For accurate and reliable gene expression analysis, normalization of gene expression data against housekeeping genes (reference or internal control genes) is required. It is known that commonly used housekeeping genes (e.g. ACTB, GAPDH, HPRT1, and B2M) vary considerably under different experimental conditions and therefore their use for normalization is limited. We performed a meta-analysis of 13,629 human gene array samples in order to identify the most stable expressed genes. Here we show novel candidate housekeeping genes (e.g. RPS13, RPL27, RPS20 and OAZ1) with enhanced stability among a multitude of different cell types and varying experimental conditions. None of the commonly used housekeeping genes were present in the top 50 of the most stable expressed genes. In addition, using 2,543 diverse mouse gene array samples we were able to confirm the enhanced stability of the candidate novel housekeeping genes in another mammalian species. Therefore, the identified novel candidate housekeeping genes seem to be the most appropriate choice for normalizing gene expression data.
de Jonge
Hendrik J M
HJ
Division of Pediatric Oncology/Hematology, Department of Pediatrics, Beatrix Children's Hospital, Groningen, The Netherlands.
Fehrmann
Rudolf S N
RS
de Bont
Eveline S J M
ES
Hofstra
Robert M W
RM
Gerbens
Frans
F
Kamps
Willem A
WA
de Vries
Elisabeth G E
EG
van der Zee
Ate G J
AG
te Meerman
Gerard J
GJ
ter Elst
Arja
A
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D020043:Genes, Essential; D006801:Humans; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D012641:Selection, Genetic
D017931:DNA Primers