Single-cell RNAseq is an emerging technology that allows the quantification of gene expression in individual cells. In plants, single-cell sequencing technology has been applied to generate root cell expression maps under many experimental conditions. DAP-seq and ATAC-seq have also been used to generate genome-scale maps of protein-DNA interactions and open chromatin regions in plants. In this protocol, we describe a multistep computational pipeline for the integration of single-cell RNAseq data with DAP-seq and ATAC-seq data to predict regulatory networks and key regulatory genes. Our approach utilizes machine learning methods including feature selection and stability selection to identify candidate regulatory genes. The network generated by this pipeline can be used to provide a putative annotation of gene regulatory modules and to identify candidate transcription factors that could play a key role in specific cell types.In this book chapter, we introduce a pipeline to mine significant biomedical entities (or bioentities) in biological networks. Our focus is on prioritizing both bioentities themselves and the associations between bioentities in order to reveal their biological functions. We will introduce three tools BEERE, WIPER, and PAGER 2.0 that can be used together for network analysis and function interpretation (1) BEERE is a network analysis tool for "Biomedical Entity Expansion, Ranking and Explorations," (2) WIPER is an entity-to-entity association ranking tool, and (3) PAGER 2.0 is a service for gene enrichment analysis.With the popularity of high-throughput transcriptomic techniques like RNAseq, models of gene regulatory networks have been important tools for understanding how genes are regulated. These transcriptomic datasets are usually assumed to reflect their associated proteins. This assumption, however, ignores post-transcriptional, translational, and post-translational regulatory mechanisms that regulate protein abundance but not transcript abundance. Here we describe a method to model cross-regulatory influences between the transcripts and proteins of a set of genes using abundance data collected from a series of transgenic experiments. The developed model can capture the effects of regulation that impacts transcription as well as regulatory mechanisms occurring after transcription. This approach uses a sparse maximum likelihood algorithm to determine relationships that influence transcript and protein abundance. An example of how to explore the network topology of this type of model is also presented. This model can be used to predict how the transcript and protein abundances will change in novel transgenic modification strategies.The cell expresses various genes in specific contexts with respect to internal and external perturbations to invoke appropriate responses. Transcription factors (TFs) orchestrate and define the expression level of genes by binding to their regulatory regions. Dysregulated expression of TFs often leads to aberrant expression changes of their target genes and is responsible for several diseases including cancers. In the last two decades, several studies experimentally identified target genes of several TFs. However, these studies are limited to a small fraction of the total TFs encoded by an organism, and only for those amenable to experimental settings. Experimental limitations lead to many computational techniques having been proposed to predict target genes of TFs. Linear modeling of gene expression is one of the most promising computational approaches, readily applicable to the thousands of expression datasets available in the public domain across diverse phenotypes. https://www.selleckchem.com/products/ecc5004-azd5004.html Linear models assume that the expression of a gene is the sum of expression of TFs regulating it. In this chapter, I introduce mathematical programming for the linear modeling of gene expression, which has certain advantages over the conventional statistical modeling approaches. It is fast, scalable to genome level and most importantly, allows mixed integer programming to tune the model outcome with prior knowledge on gene regulation.Diverse cellular phenotypes are determined by groups of transcription factors (TFs) and other regulators that influence each others' gene expression, forming transcriptional gene regulatory networks (GRNs). In many biological contexts, especially in development and associated diseases, the expression of the genes in GRNs is not static but evolves in time. Modeling the dynamics of GRN state is an important approach for understanding diverse cellular phenomena such as cell-fate specification, pluripotency and cell-fate reprogramming, oncogenesis, and tissue regeneration. In this protocol, we describe how to model GRNs using a data-driven dynamic modeling methodology, gene circuits. Gene circuits do not require knowledge of the GRN topology and connectivity but instead learn them from training data, making them very general and applicable to diverse biological contexts. We utilize the MATLAB-based gene circuit modeling software Fast Inference of Gene Regulation (FIGR) for training the model on quantitative gene expression data and simulating the GRN. We describe all the steps in the modeling life cycle, from formulating the model, training the model using FIGR, simulating the GRN, to analyzing and interpreting the model output. This protocol highlights these steps with the example of a dynamical model of the gap gene GRN involved in Drosophila segmentation and includes example MATLAB statements for each step.Gene expression data analysis and the prediction of causal relationships within gene regulatory networks (GRNs) have guided the identification of key regulatory factors and unraveled the dynamic properties of biological systems. However, drawing accurate and unbiased conclusions requires a comprehensive understanding of relevant tools, computational methods, and their workflows. The topics covered in this chapter encompass the entire workflow for GRN inference including (1) experimental design; (2) RNA sequencing data processing; (3) differentially expressed gene (DEG) selection; (4) clustering prior to inference; (5) network inference techniques; and (6) network visualization and analysis. Moreover, this chapter aims to present a workflow feasible and accessible for plant biologists without a bioinformatics or computer science background. To address this need, TuxNet, a user-friendly graphical user interface that integrates RNA sequencing data analysis with GRN inference, is chosen for the purpose of providing a detailed tutorial.