scholarly journals GeneTonic: an R/Bioconductor package for streamlining the interpretation of RNA-seq data

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Federico Marini ◽  
Annekathrin Ludt ◽  
Jan Linke ◽  
Konstantin Strauch
Keyword(s):  
Differential Expression ◽  
Differential Expression Analysis ◽  
Transcriptome Profiling ◽  
R Package ◽  
Functional Enrichment ◽  
Rna Seq ◽  
Essential Information ◽  
Wide Range ◽  
Functional Patterns ◽  
Interactive Visualizations

Abstract Background The interpretation of results from transcriptome profiling experiments via RNA sequencing (RNA-seq) can be a complex task, where the essential information is distributed among different tabular and list formats—normalized expression values, results from differential expression analysis, and results from functional enrichment analyses. A number of tools and databases are widely used for the purpose of identification of relevant functional patterns, yet often their contextualization within the data and results at hand is not straightforward, especially if these analytic components are not combined together efficiently. Results We developed the software package, which serves as a comprehensive toolkit for streamlining the interpretation of functional enrichment analyses, by fully leveraging the information of expression values in a differential expression context. is implemented in R and Shiny, leveraging packages that enable HTML-based interactive visualizations for executing drilldown tasks seamlessly, viewing the data at a level of increased detail. is integrated with the core classes of existing Bioconductor workflows, and can accept the output of many widely used tools for pathway analysis, making this approach applicable to a wide range of use cases. Users can effectively navigate interlinked components (otherwise available as flat text or spreadsheet tables), bookmark features of interest during the exploration sessions, and obtain at the end a tailored HTML report, thus combining the benefits of both interactivity and reproducibility. Conclusion is distributed as an R package in the Bioconductor project (https://bioconductor.org/packages/GeneTonic/) under the MIT license. Offering both bird’s-eye views of the components of transcriptome data analysis and the detailed inspection of single genes, individual signatures, and their relationships, aims at simplifying the process of interpretation of complex and compelling RNA-seq datasets for many researchers with different expertise profiles.

scholarly journals ideal: an R/Bioconductor package for Interactive Differential Expression Analysis

2020 ◽  
Author(s):  
Federico Marini ◽  
Jan Linke ◽  
Harald Binder
Keyword(s):  
Differential Expression ◽  
Expression Analysis ◽  
Web Application ◽  
Differential Expression Analysis ◽  
Transcriptome Profiling ◽  
Data Interpretation ◽  
R Package ◽  
Rna Seq ◽  
Fully Integrated ◽  
Bioconductor Project

AbstractBackgroundRNA sequencing (RNA-seq) is an ever increasingly popular tool for transcriptome profiling. A key point to make the best use of the available data is to provide software tools that are easy to use but still provide flexibility and transparency in the adopted methods. Despite the availability of many packages focused on detecting differential expression, a method to streamline this type of bioinformatics analysis in a comprehensive, accessible, and reproducible way is lacking.ResultsWe developed the ideal software package, which serves as a web application for interactive and reproducible RNA-seq analysis, while producing a wealth of visualizations to facilitate data interpretation. ideal is implemented in R using the Shiny framework, and is fully integrated with the existing core structures of the Bioconductor project. Users can perform the essential steps of the differential expression analysis work-flow in an assisted way, and generate a broad spectrum of publication-ready outputs, including diagnostic and summary visualizations in each module, all the way down to functional analysis. ideal also offers the possibility to seamlessly generate a full HTML report for storing and sharing results together with code for reproducibility.Conclusionideal is distributed as an R package in the Bioconductor project (http://bioconductor.org/packages/ideal/), and provides a solution for performing interactive and reproducible analyses of summarized RNA-seq expression data, empowering researchers with many different profiles (life scientists, clinicians, but also experienced bioinformaticians) to make the ideal use of the data at hand.

scholarly journals ideal: an R/Bioconductor package for interactive differential expression analysis

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Federico Marini ◽  
Jan Linke ◽  
Harald Binder
Keyword(s):  
Differential Expression ◽  
Expression Analysis ◽  
Web Application ◽  
Differential Expression Analysis ◽  
Transcriptome Profiling ◽  
Data Interpretation ◽  
R Package ◽  
Rna Seq ◽  
Bioconductor Project ◽  
Analysis Workflow

Abstract Background RNA sequencing (RNA-seq) is an ever increasingly popular tool for transcriptome profiling. A key point to make the best use of the available data is to provide software tools that are easy to use but still provide flexibility and transparency in the adopted methods. Despite the availability of many packages focused on detecting differential expression, a method to streamline this type of bioinformatics analysis in a comprehensive, accessible, and reproducible way is lacking. Results We developed the software package, which serves as a web application for interactive and reproducible RNA-seq analysis, while producing a wealth of visualizations to facilitate data interpretation. is implemented in R using the Shiny framework, and is fully integrated with the existing core structures of the Bioconductor project. Users can perform the essential steps of the differential expression analysis workflow in an assisted way, and generate a broad spectrum of publication-ready outputs, including diagnostic and summary visualizations in each module, all the way down to functional analysis. also offers the possibility to seamlessly generate a full HTML report for storing and sharing results together with code for reproducibility. Conclusion is distributed as an R package in the Bioconductor project (http://bioconductor.org/packages/ideal/), and provides a solution for performing interactive and reproducible analyses of summarized RNA-seq expression data, empowering researchers with many different profiles (life scientists, clinicians, but also experienced bioinformaticians) to make the ideal use of the data at hand.

scholarly journals Topconfects: a package for confident effect sizes in differential expression analysis provides improved usability ranking genes of interest

2018 ◽  
Author(s):  
Paul F. Harrison ◽  
Andrew D. Pattison ◽  
David R. Powell ◽  
Traude H. Beilharz
Keyword(s):  
Gene Expression ◽  
Differential Expression ◽  
Expression Analysis ◽  
Effect Size ◽  
Differential Expression Analysis ◽  
R Package ◽  
Confidence Bound ◽  
Data Set ◽  
Cancer Data ◽  
Wide Range

AbstractBackgroundA differential gene expression analysis may produce a set of significantly differentially expressed genes that is too large to easily investigate, so that a means of ranking genes by their biological interest level is desirable. The life-sciences have grappled with the abuse of p-values to rank genes for this purpose. As an alternative, a lower confidence bound on the magnitude of Log Fold Change (LFC) could be used to rank genes, but it has been unclear how to reconcile this with the need to perform False Discovery Rate (FDR) correction. The TREAT test of McCarthy and Smyth is a step in this direction, finding genes significantly exceeding a specified LFC threshold. Here we describe the use of test inversion on TREAT to present genes ranked by a confidence bound on the LFC, while still controlling FDR.ResultsTesting the Topconfects R package with simulated gene expression data shows the method outperforming current statistical approaches across a wide range of experiment sizes in the identification of genes with largest LFCs. Applying the method to a TCGA breast cancer data-set shows the method ranks some genes with large LFC higher than would traditional ranking by p-value. Importantly these two ranking methods lead to a different biological emphasis, in terms both of specific highly ranked genes and gene-set enrichment.ConclusionsThe choice of ranking method in differential expression analysis can affect the biological interpretation. The common default of ranking by p-value is implicitly by an effect size in which each gene is standardized to its own variability, rather than comparing genes on a common scale, which may not be appropriate. The Topconfects approach of presenting genes ranked by confident LFC effect size is a variation on the TREAT method with improved usability, removing the need to fine-tune a threshold parameter and removing the temptation to abuse p-values as a de-facto effect size.

scholarly journals BingleSeq: A user-friendly R package for Bulk and Single-cell RNA-Seq Data Analysis

2020 ◽  
Author(s):  
Daniel Dimitrov ◽  
Quan Gu
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
High Throughput Sequencing ◽  
Gene Annotation ◽  
Differential Expression Analysis ◽  
Transcriptome Profiling ◽  
R Package ◽  
Rna Seq ◽  
The Individual ◽  
User Friendly

AbstractRNA sequencing is a high-throughput sequencing technique considered as an indispensable research tool used in a broad range of transcriptome analysis studies. The most common application of RNA Sequencing is Differential Expression analysis and it is used to determine genetic loci with distinct expression across different conditions. On the other hand, an emerging field called single-cell RNA sequencing is used for transcriptome profiling at the individual cell level. The standard protocols for both these types of analyses include the processing of sequencing libraries and result in the generation of count matrices. An obstacle to these analyses and the acquisition of meaningful results is that both require programming expertise.BingleSeq was developed as an intuitive application that provides a user-friendly solution for the analysis of count matrices produced by both Bulk and Single-cell RNA-Seq experiments. This was achieved by building an interactive dashboard-like user interface and incorporating three state-of-the-art software packages for each type of the aforementioned analyses, alongside additional features such as key visualisation techniques, functional gene annotation analysis and rank-based consensus for differential gene analysis results, among others. As a result, BingleSeq puts the best and most widely used packages and tools for RNA-Seq analyses at the fingertips of biologists with no programming experience.

scholarly journals scRMD: imputation for single cell RNA-seq data via robust matrix decomposition

2020 ◽  
Vol 36 (10) ◽  
pp. 3156-3161 ◽  
Author(s):  
Chong Chen ◽  
Changjing Wu ◽  
Linjie Wu ◽  
Xiaochen Wang ◽  
Minghua Deng ◽  
...  
Keyword(s):  
Data Analysis ◽  
Single Cell ◽  
Differential Expression ◽  
Expression Analysis ◽  
Differential Expression Analysis ◽  
Matrix Decomposition ◽  
Transcriptome Profiling ◽  
R Package ◽  
Supplementary Information ◽  
Downstream Analysis

Abstract Motivation Single cell RNA-sequencing (scRNA-seq) technology enables whole transcriptome profiling at single cell resolution and holds great promises in many biological and medical applications. Nevertheless, scRNA-seq often fails to capture expressed genes, leading to the prominent dropout problem. These dropouts cause many problems in down-stream analysis, such as significant increase of noises, power loss in differential expression analysis and obscuring of gene-to-gene or cell-to-cell relationship. Imputation of these dropout values can be beneficial in scRNA-seq data analysis. Results In this article, we model the dropout imputation problem as robust matrix decomposition. This model has minimal assumptions and allows us to develop a computational efficient imputation method called scRMD. Extensive data analysis shows that scRMD can accurately recover the dropout values and help to improve downstream analysis such as differential expression analysis and clustering analysis. Availability and implementation The R package scRMD is available at https://github.com/XiDsLab/scRMD. Supplementary information Supplementary data are available at Bioinformatics online.

scholarly journals BingleSeq: a user-friendly R package for bulk and single-cell RNA-Seq data analysis

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e10469
Author(s):  
Daniel Dimitrov ◽  
Quan Gu
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
Differential Expression Analysis ◽  
Transcriptome Profiling ◽  
R Package ◽  
Rna Seq ◽  
Single Cell Rna Sequencing ◽  
The Individual ◽  
Visualization Techniques ◽  
User Friendly

Background RNA sequencing is an indispensable research tool used in a broad range of transcriptome analysis studies. The most common application of RNA Sequencing is differential expression analysis and it is used to determine genetic loci with distinct expression across different conditions. An emerging field called single-cell RNA sequencing is used for transcriptome profiling at the individual cell level. The standard protocols for both of these approaches include the processing of sequencing libraries and result in the generation of count matrices. An obstacle to these analyses and the acquisition of meaningful results is that they require programing expertise. Although some effort has been directed toward the development of user-friendly RNA-Seq analysis analysis tools, few have the flexibility to explore both Bulk and single-cell RNA sequencing. Implementation BingleSeq was developed as an intuitive application that provides a user-friendly solution for the analysis of count matrices produced by both Bulk and Single-cell RNA-Seq experiments. This was achieved by building an interactive dashboard-like user interface which incorporates three state-of-the-art software packages for each type of the aforementioned analyses. Furthermore, BingleSeq includes additional features such as visualization techniques, extensive functional annotation analysis and rank-based consensus for differential gene analysis results. As a result, BingleSeq puts some of the best reviewed and most widely used packages and tools for RNA-Seq analyses at the fingertips of biologists with no programing experience. Availability BingleSeq is as an easy-to-install R package available on GitHub at https://github.com/dbdimitrov/BingleSeq/.

scholarly journals Polyester: simulating RNA-seq datasets with differential transcript expression

2014 ◽  
Author(s):  
Alyssa C Frazee ◽  
Andrew E Jaffe ◽  
Ben Langmead ◽  
Jeffrey Leek
Keyword(s):  
Differential Expression ◽  
Rate Control ◽  
Differential Expression Analysis ◽  
R Package ◽  
Experimental Designs ◽  
Rna Seq ◽  
Reasonable Approximation ◽  
Methods Development ◽  
Error Rate Control ◽  
Expression Status

Motivation: Statistical methods development for differential expression analysis of RNA sequencing (RNA-seq) requires software tools to assess accuracy and error rate control. Since true differential expression status is often unknown in experimental datasets, artificially-constructed datasets must be utilized, either by generating costly spike-in experiments or by simulating RNA-seq data. Results: Polyester is an R package designed to simulate RNA-seq data, beginning with an experimental design and ending with collections of RNA-seq reads. Its main advantage is the ability to simulate reads indicating isoform-level differential expression across biological replicates for a variety of experimental designs. Data generated by Polyester is a reasonable approximation to real RNA-seq data and standard differential expression workflows can recover differential expression set in the simulation by the user. Availability and Implementation: Polyester is freely available from Bioconductor (http://bioconductor.org/).

scholarly journals rmRNAseq: differential expression analysis for repeated-measures RNA-seq data

2020 ◽  
Vol 36 (16) ◽  
pp. 4432-4439 ◽  
Author(s):  
Yet Nguyen ◽  
Dan Nettleton
Keyword(s):  
Differential Expression ◽  
Expression Analysis ◽  
Repeated Measures ◽  
Differential Expression Analysis ◽  
Parametric Bootstrap ◽  
R Package ◽  
Experimental Unit ◽  
Multiple Time ◽  
Rna Seq ◽  
Sequencing Technologies

Abstract Motivation With the reduction in price of next-generation sequencing technologies, gene expression profiling using RNA-seq has increased the scope of sequencing experiments to include more complex designs, such as designs involving repeated measures. In such designs, RNA samples are extracted from each experimental unit at multiple time points. The read counts that result from RNA sequencing of the samples extracted from the same experimental unit tend to be temporally correlated. Although there are many methods for RNA-seq differential expression analysis, existing methods do not properly account for within-unit correlations that arise in repeated-measures designs. Results We address this shortcoming by using normalized log-transformed counts and associated precision weights in a general linear model pipeline with continuous autoregressive structure to account for the correlation among observations within each experimental unit. We then utilize parametric bootstrap to conduct differential expression inference. Simulation studies show the advantages of our method over alternatives that do not account for the correlation among observations within experimental units. Availability and implementation We provide an R package rmRNAseq implementing our proposed method (function TC_CAR1) at https://cran.r-project.org/web/packages/rmRNAseq/index.html. Reproducible R codes for data analysis and simulation are available at https://github.com/ntyet/rmRNAseq/tree/master/simulation.

scholarly journals Best practices on the differential expression analysis of multi-species RNA-seq

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Matthew Chung ◽  
Vincent M. Bruno ◽  
David A. Rasko ◽  
Christina A. Cuomo ◽  
José F. Muñoz ◽  
...  
Keyword(s):  
Best Practices ◽  
Differential Expression ◽  
Expression Analysis ◽  
Differential Expression Analysis ◽  
Single Species ◽  
Rna Seq ◽  
Species Analysis ◽  
Differential Gene ◽  
Multiple Species ◽  
Downstream Analysis

AbstractAdvances in transcriptome sequencing allow for simultaneous interrogation of differentially expressed genes from multiple species originating from a single RNA sample, termed dual or multi-species transcriptomics. Compared to single-species differential expression analysis, the design of multi-species differential expression experiments must account for the relative abundances of each organism of interest within the sample, often requiring enrichment methods and yielding differences in total read counts across samples. The analysis of multi-species transcriptomics datasets requires modifications to the alignment, quantification, and downstream analysis steps compared to the single-species analysis pipelines. We describe best practices for multi-species transcriptomics and differential gene expression.

scholarly journals MUREN: a robust and multi-reference approach of RNA-seq transcript normalization

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Yance Feng ◽  
Lei M. Li
Keyword(s):  
Biological Significance ◽  
Housekeeping Genes ◽  
R Package ◽  
Data Sets ◽  
Statistical Regression ◽  
Rna Seq ◽  
Least Trimmed Squares ◽  
Standard Data ◽  
Wide Range ◽  
Multiple References

Abstract Background Normalization of RNA-seq data aims at identifying biological expression differentiation between samples by removing the effects of unwanted confounding factors. Explicitly or implicitly, the justification of normalization requires a set of housekeeping genes. However, the existence of housekeeping genes common for a very large collection of samples, especially under a wide range of conditions, is questionable. Results We propose to carry out pairwise normalization with respect to multiple references, selected from representative samples. Then the pairwise intermediates are integrated based on a linear model that adjusts the reference effects. Motivated by the notion of housekeeping genes and their statistical counterparts, we adopt the robust least trimmed squares regression in pairwise normalization. The proposed method (MUREN) is compared with other existing tools on some standard data sets. The goodness of normalization emphasizes on preserving possible asymmetric differentiation, whose biological significance is exemplified by a single cell data of cell cycle. MUREN is implemented as an R package. The code under license GPL-3 is available on the github platform: github.com/hippo-yf/MUREN and on the conda platform: anaconda.org/hippo-yf/r-muren. Conclusions MUREN performs the RNA-seq normalization using a two-step statistical regression induced from a general principle. We propose that the densities of pairwise differentiations are used to evaluate the goodness of normalization. MUREN adjusts the mode of differentiation toward zero while preserving the skewness due to biological asymmetric differentiation. Moreover, by robustly integrating pre-normalized counts with respect to multiple references, MUREN is immune to individual outlier samples.

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