EndoGeneAnalyzer: A tool for selection and validation of reference genes

PLoS One. 2024 Apr 3;19(4):e0299993. doi: 10.1371/journal.pone.0299993. eCollection 2024.

Abstract

The selection of proper reference genes is critical for accurate gene expression analysis in all fields of biological and medical research, mainly because there are many distinctions between different tissues and specimens. Given this variability, even in known classic reference genes, demands of a comprehensive analysis platform is needed to identify the most suitable genes for each study. For this purpose, we present an analysis tool for assisting in decision-making in the analysis of reverse transcription-quantitative polymerase chain reaction (RT-qPCR) data. EndoGeneAnalyzer, an open-source web tool for reference gene analysis in RT-qPCR studies, was used to compare the groups/conditions under investigation. This interactive application offers an easy-to-use interface that allows efficient exploration of datasets. Through statistical and stability analyses, EndoGeneAnalyzer assists in the select of the most appropriate reference gene or set of genes for each condition. It also allows researchers to identify and remove unwanted outliers. Moreover, EndoGeneAnalyzer provides a graphical interface to compare the evaluated groups, providing a visually informative differential analysis.

MeSH terms

  • Gene Expression Profiling*
  • Real-Time Polymerase Chain Reaction
  • Reference Standards
  • Reverse Transcriptase Polymerase Chain Reaction

Grants and funding

This study was supported by Brazilian funding agencies: Coordination for the Improvement of Higher Education Personnel (CAPES; to E.B.T), the National Council of Technological and Scientific Development (CNPq grant number [404213/2021-9 to CAM-N; Productivity in Research PQ scholarships to P.P.A, A.S.K., and CAM-N]), and the Cearense Foundation of Scientific and Technological Support (FUNCAP grant number [P20-0171-00078.01.00/20 to CAM-N]); we also thank PROPESP/UFPA for the publication payment. There was no additional external funding received for this study.