In search of suitable reference genes for gene expression studies of human renal cell carcinoma by real-time PCR

Jung, Monika; Ramankulov, Azizbek; Roigas, Jan; Johannsen, Manfred; Ringsdorf, Martin; Kristiansen, Glen; Jung, Klaus

doi:10.1186/1471-2199-8-47

Cited by 143 publications

(130 citation statements)

References 40 publications

Supporting

Mentioning

125

Contrasting

Order By: Relevance

“…A number of methods, such as GeNorm, allow identification of suitable reference genes [21,24,25]. Since these tools have become available, many papers have highlighted the need to identify suitable reference genes in various biological systems, whether they compare treated versus untreated samples or examine developmentally regulated gene expression [32][33][34][35]. However, suitable reference genes that are stably expressed in various cell types of the developing male and female gonads have not yet been identified.…”

Section: Discussionmentioning

confidence: 99%

Normalizing Gene Expression Levels in Mouse Fetal Germ Cells1

Bergen

Miles

Sinclair

et al. 2009

Biology of Reproduction

View full text Add to dashboard Cite

Real-time PCR has become a popular method to analyze transcription of genes that are developmentally regulated during organogenesis of the testes and ovaries. However, the heterogenous cell populations and commitment to strikingly different developmental pathways of the germ and somatic cells in these organs complicate analysis of this process. The selection of suitable reference genes for quantifying gene expression in this system is essential, but to date it has not been sufficiently addressed. To rectify this problem, we have used fluorescenceactivated cell sorting to purify germ cells from mouse fetal testes and ovaries and examined 16 common housekeeping genes for their suitability as reference genes. In pure populations of germ cells isolated from Embryonic Day 12.5 (E12.5) to E15.5 male and female gonads, Mapk1 and Sdha were identified as the most stable reference genes. Analysis of the heterogenous fraction of gonadal somatic cells revealed that Canx and Top1 were stable in both sexes, whereas a comparative analysis of germ and somatic cell populations identified Canx and Mapk1 as suitable reference genes through these developmental stages. Application of these reference genes to quantification of gene expression in developing gonads revealed that past assays, which employed nonverified reference genes, have in some cases provided misleading gene expression profiles. This study has identified suitable reference genes to directly compare expression profiles of genes expressed in germ and somatic cells of male and female fetal gonads. Application of these reference genes to expression analysis in fetal germ and somatic cells provides a more accurate system in which to profile gene expression in these tissues.

show abstract

Section: Discussionmentioning

confidence: 99%

Normalizing Gene Expression Levels in Mouse Fetal Germ Cells1

Bergen

Miles

Sinclair

et al. 2009

Biology of Reproduction

View full text Add to dashboard Cite

show abstract

“…The primers were designed, using the Primer3 (http://frodo.wi.mit.edu/primer3/) and IDT (http://eu.idtdna.com/site) programs (Table 1). TBP was used as a reference gene [9].…”

mentioning

confidence: 99%

PPM1M and PRICKLE2 are potential tumor suppressor genes in human clear-cell renal cell carcinoma

et al. 2014

View full text Add to dashboard Cite

show abstract

“…Such "standard" reference genes such as GAPDH (glyceraldehyde 3-phosphate dehydrogenase), ACTB (beta-actin), RN18S1, and RN28S1 (ribosomal RNA genes), RPLP0 (a ribosomal protein), MHC I (major histocompatibility complex I), B2M (beta-2 microglobulin), and GUSB (glucuronidase) have been previously used to study human and animal tissues (Huggett et al, 2005;Hoogewijs et al, 2008). However, according to many studies, even the "traditional" housekeeping genes might be differentially expressed in some cases (Jung et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

A new reliable reference gene UBA52 for quantitative real-time polymerase chain reaction studies in pyloric cecal tissues of the starfish Asterias rubens

Садритдинова¹,

Dmitriev²,

Снежкина³

et al. 2014

Genet. Mol. Res.

View full text Add to dashboard Cite

ABSTRACT. The starfish Asterias rubens is one of the most abundant echinoderm species in the White, Barents, North, and Baltic Seas. This species is an important component of marine ecosystems and a model object for certain biological studies, in particular those requiring quantitative estimation of gene expression. As a rule, expression at the transcriptional level is estimated by real-time qPCR using the ΔΔCt method, which allows the comparison of the copy number of target gene transcripts in samples with unknown mRNA/cDNA concentration. Application of this method requires normalization of the results relative to genes with stable expression levels (reference genes). The identification of reference genes is still a challenging task since data of this kind are missing for certain taxa, whereas the use of "standard" endogenous control genes without additional tests might lead to erroneous conclusions. We performed a preliminary analysis of the expression of many housekeeping genes in the pyloric ceca of A. rubens by high-throughput sequencing under normal and heat shock conditions. For one of them, the ubiquitin gene UBA52, low variation of expression (not greater than 2-fold) was shown using real-time qPCR. Tissues of pyloric ceca of normal adults and underyearlings and of adults after heat shock were used. The data obtained suggest that the UBA52 gene may be used as reference for normalization of gene expression at the mRNA level in the starfish A. rubens and probably in closely related species.

show abstract

In search of suitable reference genes for gene expression studies of human renal cell carcinoma by real-time PCR

Cited by 143 publications

References 40 publications

Normalizing Gene Expression Levels in Mouse Fetal Germ Cells1

Normalizing Gene Expression Levels in Mouse Fetal Germ Cells1

PPM1M and PRICKLE2 are potential tumor suppressor genes in human clear-cell renal cell carcinoma

A new reliable reference gene UBA52 for quantitative real-time polymerase chain reaction studies in pyloric cecal tissues of the starfish Asterias rubens

Contact Info

Product

Resources

About