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MS2 Phage Ribonucleoproteins as Exogenous Internal Control for RT-qPCR Data Normalization in Gene Expression Study of Developing Rat Brain

L. A. Fedoseeva1*, O. B. Shevelev1, N. G. Kolosova1, and G. M. Dymshits2

1Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, pr. Akademika Lavrent’eva 10, 630090 Novosibirsk, Russia; E-mail: fedoseeva@bionet.nsc.ru

2Novosibirsk State University, ul. Pirogova 2, 630090 Novosibirsk, Russia

* To whom correspondence should be addressed.

Received January 24, 2014; Revision received March 18, 2014
The most popular strategy for normalization of RT-qPCR data involves presenting them in comparison with expression of “housekeeping” genes. However, the required stable expression of the control genes is not always achievable. As an alternative, we used ribonucleoprotein phage particles as an exogenous internal control and demonstrated that this type of normalization provides a simple and reliable method for quantification in RT-qPCR experiments. Using phage-based normalization, we analyzed mRNA levels of three popular housekeeping genes coding β-actin, glyceraldehyde-3-phosphate dehydrogenase, and ribosomal protein L30 and showed high variability in their expression patterns during rat brain development, indicating that they should not be used as controls in gene expression studies of the developing brain either individually or in combination. Using phage-based controls, we showed interstrain differences and age-related changes in the expression of genes involved in proteoglycan biosynthesis and degradation in developing brain of senescence-accelerated OXYS rats and control Wistar rats.
KEY WORDS: PCR data normalization, reference genes, housekeeping genes, exogenous internal control, ribonucleoprotein particles, senescence-accelerated OXYS rats

DOI: 10.1134/S0006297914070128