MS2 phage ribonucleoproteins as exogenous internal control for RT-qPCR data normalization in gene expression study of developing rat brain
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 exogen...
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| Veröffentlicht in: | Biochemistry (Moscow) 2014-07, Vol.79 (7), p.706-716 |
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| creator | Fedoseeva, L. A. Shevelev, O. B. Kolosova, N. G. Dymshits, G. M. |
| description | 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 senescenceaccelerated OXYS rats and control Wistar rats. |
| doi_str_mv | 10.1134/S0006297914070128 |
| format | Article |
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A. ; Shevelev, O. B. ; Kolosova, N. G. ; Dymshits, G. M.</creator><creatorcontrib>Fedoseeva, L. A. ; Shevelev, O. B. ; Kolosova, N. G. ; Dymshits, G. M.</creatorcontrib><description>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. 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A.</creatorcontrib><creatorcontrib>Shevelev, O. B.</creatorcontrib><creatorcontrib>Kolosova, N. G.</creatorcontrib><creatorcontrib>Dymshits, G. M.</creatorcontrib><title>MS2 phage ribonucleoproteins as exogenous internal control for RT-qPCR data normalization in gene expression study of developing rat brain</title><title>Biochemistry (Moscow)</title><addtitle>Biochemistry Moscow</addtitle><addtitle>Biochemistry (Mosc)</addtitle><description>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 senescenceaccelerated OXYS rats and control Wistar rats.</description><subject>Animals</subject><subject>Biochemistry</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Bioorganic Chemistry</subject><subject>Biosynthesis</subject><subject>Brain</subject><subject>Brain - growth & development</subject><subject>Brain - metabolism</subject><subject>Brain research</subject><subject>Gene expression</subject><subject>Gene Expression Profiling - standards</subject><subject>Genes, Essential</subject><subject>Genetic aspects</subject><subject>Levivirus</subject><subject>Life Sciences</subject><subject>Male</subject><subject>Microbiology</subject><subject>Nucleoproteins</subject><subject>Physiological aspects</subject><subject>Polymerase chain reaction</subject><subject>Protein research</subject><subject>Proteins</subject><subject>Rats, Wistar</subject><subject>Real-Time Polymerase Chain Reaction - standards</subject><subject>Reference Standards</subject><subject>Reverse Transcriptase Polymerase Chain Reaction - standards</subject><subject>Ribonucleoproteins - genetics</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>Rodents</subject><subject>Transcriptome</subject><subject>Viral Proteins - genetics</subject><issn>0006-2979</issn><issn>1608-3040</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqFkstu1DAUhi0EosPAA7BBltiwSfEtibOsRtykIlBb1pFjHwdXiT21HUR5BJ4ahymo3IS8sHzO9__ybx-EHlNyTCkXz88JIQ3r2o4K0hLK5B20oQ2RFSeC3EWbtV2t_SP0IKXLcmSk4_fREaspkZzzDfr69pzh_Uc1Ao5uCH7RE4R9DBmcT1glDJ_DCD4sCTufIXo1YR18jmHCNkR8dlFdvd-dYaOywj7EWU3ui8ou-MLjooTisI-Q0lpKeTHXOFhs4BNMYe_8iKPKeIjK-YfonlVTgkc3-xZ9ePniYve6On336s3u5LTSomW50oOWqgYlBqG1NBZko00L1khBu0GITrFGEmNI3cDAqZSs5G4tty0FYtqGb9Gzg2-JebVAyv3skoZpUh5Kzp42DRGUCE7_j9Y1k6Rjzer69Df0Mizrc32nBOU1F7eoUU3QO29Djkqvpv0Jl4KRti3fskXHf6HKMjC78vpgXan_IqAHgY4hpQi230c3q3jdU9Kvo9L_MSpF8-Tmwsswg_mp-DEbBWAHIJWWHyHeSvRP128QXMeS</recordid><startdate>20140701</startdate><enddate>20140701</enddate><creator>Fedoseeva, L. 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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. 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| subjects | Animals Biochemistry Biomedical and Life Sciences Biomedicine Bioorganic Chemistry Biosynthesis Brain Brain - growth & development Brain - metabolism Brain research Gene expression Gene Expression Profiling - standards Genes, Essential Genetic aspects Levivirus Life Sciences Male Microbiology Nucleoproteins Physiological aspects Polymerase chain reaction Protein research Proteins Rats, Wistar Real-Time Polymerase Chain Reaction - standards Reference Standards Reverse Transcriptase Polymerase Chain Reaction - standards Ribonucleoproteins - genetics RNA, Messenger - genetics RNA, Messenger - metabolism Rodents Transcriptome Viral Proteins - genetics |
| title | MS2 phage ribonucleoproteins as exogenous internal control for RT-qPCR data normalization in gene expression study of developing rat brain |
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