This volume describes a variety of protocols that will allow the readers to study different aspects of transcriptional and posttranscriptional gene expression regulation in eukaryotic cells. Chapters focus on the latest use of CRISPRi and RNAi technologies for studying various aspects of transcriptional and posttranscriptional regulation and tools to navigate protocols on key bioinformatics. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls
Authoritative and cutting-edge, Eukaryotic Transcription and Post-Transcription Gene Expression Regulation aims to ensure successful results in the further study of this vital field.
Table of Contents: Fluorescence Reporter-Based Genome-Wide RNA Interference Screening to Identify Alternative Splicing Regulators.- Tandem Affinity Purification Approach Coupled To Mass Spectrometry to Identify Post-translational Modifications of Histones Associated with Chromatin-binding Proteins.- Efficient Preparation of High-Complexity ChIP-Seq Profiles from Early
Xenopus Embryos.- Systematic Discovery Of Chromatin-bound Protein Complexes from ChIP-seq Datasets.- Assay for Transposase-Accessible Chromatin with high throughput sequencing (ATAC-seq) Protocol for Zebrafish Embryos.- Establishment of time- and cell-specific RNAi in
Caenorhabditis elegans.- Cell-penetrating Peptide-mediated Delivery of Cas9 Protein and Guide RNA for Genome Editing.- Epigenetic Analysis of Endocrine Cell Subtypes from Human Pancreatic Islets.- eIF3 Regulation of Protein Synthesis, Tumorigenesis, and Therapeutic Response.- High Resolution Gene Expression Profiling of RNA Synthesis, Processing, and Decay by Metabolic Labeling of Newly Transcribed RNA using 4-thiouridine.- Accurate Detection of Differential Expression and Splicing using Low-level Features.- Profiling Changes in Histone Post-translational Modifications by Top-Down Mass Spectrometry.- Determining if an mRNA is a Substrate of Nonsense-mediated mRNA Decay in Saccharomyces cerevisiae.- Optimizing In Vitro Pre-mRNA 3' Cleavage Efficiency: Reconstitution from Anion-Exchange Separated HeLa Cleavage Factors and from Adherent HeLa Cell Nuclear Extract.- Unbiased Interrogation of 3D Genome Topology using Chromosome Conformation Capture Coupled to High-throughput Sequencing (3C-Seq).- Using an Inducible CRISPR-Cas9-KRAB Effector System to Dissect Transcriptional Regulation in Human Embryonic Stem Cells.-
In vitro assay to Study Histone Ubiquitination during Transcriptional Regulation.- Determination of Alternate Splicing Events using Transcriptome Arrays.- Large-scale RNA Interference Screening to Identify Transcriptional Regulators of a Tumor Suppressor.- GeneTranscriptional Analysis-based Integrative Genomics Approach to Identify Tumor Promoting Metabolic Genes.
