RT Journal Article
SR Electronic
T1 Transcriptomic-based selection of reference genes for quantitative real-time PCR in an insect endosymbiotic model
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2023.01.31.526490
DO 10.1101/2023.01.31.526490
A1 Vallier, Agnès
A1 Dell’Aglio, Elisa
A1 Ferrarini, Mariana Galvão
A1 Hurtado, Ophélie
A1 Monégat, Carole Vincent
A1 Heddi, Abdelaziz
A1 Rebollo, Rita
A1 Zaidman-Rémy, Anna
YR 2023
UL http://biorxiv.org/content/early/2023/01/31/2023.01.31.526490.abstract
AB Reference genes are a fundamental tool for analyses of gene expression by real-time quantitative PCR (qRT-PCR), in that they ensure the correct comparison between conditions, stages, or treatments. Because of this, selection of appropriate genes to use as references is crucial for proper application of the technique. Nevertheless, efforts to find appropriate, stably expressed transcripts are still lacking, in particular in the field of insect science. Here, we took advantage of a massive transcriptomic high-throughput analysis of various developmental stages of the gut and associated-bacteriomes of the cereal weevil Sitophilus oryzae and identified a subset of stably expressed genes with the potential to be used as housekeeping genes from the larva to the adult stage. We employed several normalization techniques to select the most suitable genes among our subset. Our final selection includes three genes - TAO, YTH3 and PP12A - which can also be used to compare transcript abundance at various developmental stages in symbiotic insects, and in insects devoid of endosymbionts (aposymbiotic). Since they are well conserved, these genes have the potential to be useful for many other insect species. This work confirms the interest in using large-scale, unbiased methods for reference gene selection.Competing Interest StatementThe authors have declared no competing interest.