Essential Genes and Genomes: Methods and Protocols [Hardcover]

This book provides state-of-the-art information on gene essentiality screenings in a wide variety of organisms, i.e. screening for protein-coding genes and other genomic elements that are required by an organism to survive under specific conditions. With a focus on the two techniques that have revolutionized the field, the collection begins with chapters employing CRISPR/Cas9-based approaches followed by Tn-seq-based approaches, but later chapters also delve into other techniques for exploring essential genes, such as bioinformatics methods. Written for the highly successful Methods in Molecular Biology series, 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. A Method to Map Gene Essentiality of Human Pluripotent Stem Cells by Genome-Scale CRISPR Screens with Inducible Cas9.- CRISPR/Cas9 Screening to Identify Conditionally Essential Genes in Human Cell Lines.- Target Identification of Small Molecules Using Large-Scale CRISPR-Cas Mutagenesis Scanning of Essential Genes.- Design and Generation of a CRISPR Interference System for Genetic Repression and Essential Gene Analysis in the Fungal Pathogen Candida albicans.- Identification of Essential Genes Using Sequential CRISPR and siRNA Screens.- A Simple Method that Combines CRISPR and AID to Quickly Generate Conditional Knockouts for Essential Genes in Various Vertebrate Cell Lines.- New Method for Genome-Scale Functional Genomics Study in Bacteria with Superior Performance: CRISPR Interference Screen.- A CRISPR-Based Method for Constructing Conditional Mutations l¾