Background 

RNA-seq is a widely adopted affordable method for large scale gene expression profiling. However, user-friendly and versatile tools for wet-lab biologists to analyse RNA-seq data beyond standard analyses such as differential expression, are rare. Especially, the analysis of time-series data is difficult for wet-lab biologists lacking advanced computational training. Furthermore, most meta-analysis tools are tailored for model organisms and not easily adaptable to other species.

Results

With RNfuzzyApp, we provide a user-friendly, web-based R-shiny app for differential expression analysis, as well as time-series analysis of RNA-seq data. RNfuzzyApp offers several methods for normalization and differential expression analysis of RNA-seq data, providing easy-to-use toolboxes, interactive plots and downloadable results. For time-series analysis, RNfuzzyApp presents the first web-based, automated pipeline for soft clustering with the Mfuzz R package, including methods to aid in cluster number selection, Mfuzz loop computations, cluster overlap analysis, as well as cluster enrichments.

Conclusion

RNfuzzyApp is an intuitive, easy to use and interactive R shiny app for RNA-seq differential expression and time-series analysis, offering a rich selection of interactive plots, providing a quick overview of raw data and generating rapid analysis results. Furthermore, its orthology assignment, enrichment analysis, as well as ID conversion functions are accessible to non-model organisms.

Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt: mean values calculated from raw reads of replicates, downloaded from gene expression omnibus (dataset GSE143430 https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE143430).

Haering_etal_extendedDatatable_1a_Tabulamurissenis_3vs12m_DEA.txt: Tabula muris senis limb muscle data (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE132040) from 3, 12 and 27month males, processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_1b_Tabulamurissenis_3vs27m_DEA.txt: Tabula muris senis limb muscle data (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE132040) from 3, 12 and 27month males, processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_1c_Tabulamurissenis_12vs27m_DEA.txt: Tabula muris senis limb muscle data (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE132040) from 3, 12 and 27month males, processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_1d_Tabulamurissenis_3vs12m_gpofiler.txt: Tabula muris senis limb muscle data (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE132040) from 3, 12 and 27month males, processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_1e_Tabulamurissenis_3vs27m_gpofiler.txt: Tabula muris senis limb muscle data (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE132040) from 3, 12 and 27month males, processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_1f_Tabulamurissenis_12vs27m_gpofiler.txt: Tabula muris senis limb muscle data (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE132040) from 3, 12 and 27month males, processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_2a_Tabulamurissenis_cluster1_gpofiler.txt: Tabula muris senis limb muscle data (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE132040) from 3, 12 and 27month males, processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_2b_Tabulamurissenis_cluster2_gpofiler.txt: Tabula muris senis limb muscle data (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE132040) from 3, 12 and 27month males, processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_2c_Tabulamurissenis_cluster3_gpofiler.txt: Tabula muris senis limb muscle data (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE132040) from 3, 12 and 27month males, processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_2d_Tabulamurissenis_cluster4_gpofiler.txt: Tabula muris senis limb muscle data (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE132040) from 3, 12 and 27month males, processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_2e_Tabulamurissenis_cluster5_gpofiler.txt: Tabula muris senis limb muscle data (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE132040) from 3, 12 and 27month males, processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_3a_DmLeg_cluster1_gpofiler.txt: Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_3b_DmLeg_cluster2_gpofiler.txt: Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_3c_DmLeg_cluster3_gpofiler.txt: Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_3d_DmLeg_cluster4_gpofiler.txt: Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_3e_DmLeg_cluster5_gpofiler.txt: Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_3f_DmLeg_cluster6_gpofiler.txt: Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_3g_DmLeg_cluster7_gpofiler.txt: Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_3h_DmLeg_cluster8_gpofiler.txt: Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_3i_DmLeg_cluster9_gpofiler.txt: Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_3j_DmLeg_cluster10_gpofiler.txt: Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_3k_DmLeg_cluster11_gpofiler.txt: Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Haering_etal_extendedDatatable_3l_DmLeg_cluster12_gpofiler.txt: Haering_etal_extendedData_DmdevLeg_GSE143430_mean.txt processed with RNfuzzyApp (https://gitlab.com/habermann_lab/rna-seq-analysis-app)

Details on data processing and analysis can be found in the associated article.