Repository of Research and Investigative Information

Repository of Research and Investigative Information

دانشگاه علوم پزشکی و خدمات بهداشتی درمانی زنجان

Selection of suitable reference genes for analysis of salivary transcriptome in non-syndromic autistic male children

(2016) Selection of suitable reference genes for analysis of salivary transcriptome in non-syndromic autistic male children. International journal of molecular sciences.

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Abstract

Childhood autism is a severe form of complex genetically heterogeneous and behaviorally defined set of neurodevelopmental diseases, collectively termed as autism spectrum disorders (ASD). Reverse transcriptase quantitative real-time PCR (RT-qPCR) is a highly sensitive technique for transcriptome analysis, and it has been frequently used in ASD gene expression studies. However, normalization to stably expressed reference gene(s) is necessary to validate any alteration reported at the mRNA level for target genes. The main goal of the present study was to find the most stable reference genes in the salivary transcriptome for RT-qPCR analysis in non-syndromic male childhood autism. Saliva samples were obtained from nine drug naïve non-syndromic male children with autism and also sex-, age-, and location-matched healthy controls using the RNA-stabilizer kit from DNA Genotek. A systematic two-phased measurement of whole saliva mRNA levels for eight common housekeeping genes (HKGs) was carried out by RT-qPCR, and the stability of expression for each candidate gene was analyzed using two specialized algorithms, geNorm and NormFinder, in parallel. Our analysis shows that while the frequently used HKG ACTB is not a suitable reference gene, the combination of GAPDH and YWHAZ could be recommended for normalization of RT-qPCR analysis of salivary transcriptome in non-syndromic autistic male children. © 2016 by the authors; licensee MDPI, Basel, Switzerland.

Item Type: Article
Keywords: Childhood autism GeNorm Housekeeping genes (HKGs) Non-syndromic NormFinder Reference gene Reverse transcriptase quantitative real-time PCR (RT-qPCR) Saliva Stability of expression Transcriptome flavoprotein glyceraldehyde 3 phosphate dehydrogenase messenger RNA ribosome protein RNA 18S transferrin receptor ubiquitin C Article autism child clinical article clinical trial controlled study DNA synthesis gene expression genetic algorithm genetic analysis genetic selection housekeeping gene human male reverse transcription polymerase chain reaction RNA isolation stable expression gene expression profiling gene expression regulation genetics metabolism real time polymerase chain reaction Autistic Disorder Humans Real-Time Polymerase Chain Reaction RNA, Ribosomal, 18S
Journal or Publication Title: International journal of molecular sciences
Abstract and Indexing: ISI, Pubmed, Scopus
Quartile : Q2
Volume: 17
Number: 10
Identification Number: https://doi.org/10.3390/ijms17101711
Depositing User: خانم مریم زرقانی
URI: http://repository.zums.ac.ir/id/eprint/737

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