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Full Description
This third edition expands on the previous editions by providing updated protocols and current methods that cover recent breakthroughs in Drosophila research. Chapters guide readers through FlyBase, versatile gene expression systems, analysis of microRNA function, single-cell transcriptome data and metabolism, recent applications of CRISPR for precise genome editing, transcriptional activation and cell lineage tracing, protein inhibition and tagging, optogenetic and optochemical control of tissue mechanics, AFM measurements, sample preparation for electron microscopy, live imaging of different tissues and organs, and quantitative image analysis. Written in the format of the highly successful Methods in Molecular Biology series, chapters include an introduction to their topic, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls
Authoritative and cutting-edge, Drosophila: Methods and Protocols, Third Edition serves as a useful and practical guide to new researchers and experts using Drosophila as a model system.
Chapter 7 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
Contents
Using FlyBase, a database of Drosophila genes and genetics.- The Q-System: A versatile repressible binary expression system.- Resources and methods for the analysis of microRNA function in Drosophila.- Analysis of single-cell transcriptome data in Drosophila.- Prime editing for precise genome engineering in Drosophila.- CRISPR/Cas9 mediated precise and efficient genome editing in Drosophila.- Tissue-specific CRISPR-Cas9 screening in Drosophila.- CRISPR-based transcriptional activation in Drosophila.- Tracing and manipulating Drosophila cell lineages based on CRISPR: CaSSA and CLADES.- Studying protein function using nanobodies and other protein binders in Drosophila.- Anchor Away: A system for fast inhibition of proteins in Drosophila.- Tagging Drosophila proteins with genetically-encoded fluorophores.- Optogenetic methods to control tissue mechanicsin Drosophila.- Optochemical control of cell contractility in Drosophila embryos.- Stiffness measurement of Drosophila egg chambers by atomic force microscopy.- Cultivation and live imaging of Drosophila imaginal discs.- Sample preparation and imaging of the pupal Drosophila abdominal epidermis.- FlyClear: A tissue clearing technique for high-resolution microscopy of Drosophila.- Preparation of Drosophila tissues and organs for transmission electron microscopy.- Segmentation and quantitative analysis of epithelial tissues.- Genetically-encoded sensors to study metabolism in Drosophila.
