Drosophila: Methods and Protocols

ISBN-13: 9781493963690 / Angielski / Twarda / 2016 / 355 str.

Christian Dahmann

Opis

This second edition volume expands on the previous edition by presenting updated protocols for several of the techniques described in the first edition of Drosophila: Methods and Protocols and current methods that cover recent breakthroughs in Drosophila research. The book begins with a description of FlyBase--a database of genes and genomes--followed by the presentation of systems for versatile gene expression in the fly. The first few chapters in this book detail gene knockdown and editing, including CRISPR-Cas9 and protein knockdown. The next few chapters are devoted to methods describing live imaging of different tissues and organs, followed by chapters on how to quantify image data and how to probe tissue mechanics by laser ablation. The next two chapters provide methods for analyzing transcription followed by protocols to study growth, metabolism, ageing, and behavior in Drosophila. This volume concludes with chapters on electrophysiological recordings and methods to establish cell lines. 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.

Cutting-edge and thorough, Drosophila: Methods and Protocols, Second Edition is a valuable source of hands-on protocols and reviews for molecular, cell, and developmental biologists using Drosophila as model systems in their work.

Spis treści

1.      Using FlyBase: A Database of Drosophila Genes and Genomes
Steven J. Marygold, Madeline A. Crosby, Joshua L. Goodman, and The FlyBase Consortium

2.      The GAL4 System: A Versatile System for Manipulation and Analysis of Gene Expression
Elizabeth E. Caygill and Andrea H. Brand

3.      The Q-System: A Versatile Expression System for Drosophila
Olena Riabinina and Christopher J. Potter

4.      Analysis of MicroRNA Function in Drosophila
Geetanjali Chawla, Arthur Luhur, and Nicholas Sokol

5.      Methods for High-Throughput RNAi screening in Drosophila Cells
Maximilian Billmann and Michael Boutros

6.      A Guide to Genome-Wide In Vivo RNAi Applications in Drosophila
Aynur Kaya-Çopur and Frank Schnorrer

7.      Creating Heritable Mutations in Drosophila with CRISPR-Cas9
Fillip Port and Simon L. Bullock

8.      Performing Chromophore-Assisted Laser Inactivation in Drosophila Embryos Using GFP
Anne Pélissier-Monier, Bénédicte Sanson, and Bruno Monier

9.      deGradFP: A System to Knockdown GFP-Tagged Proteins
Emmanuel Caussinus and Markus Affolter

10.  Sample Preparation and Mounting of Drosophila Embryos for Multi-View Light Sheet Microscopy
Christopher Schmied and Pavel Tomancak

11.  Cultivation and Live-Imaging of Drosophila Imaginal DiscsSimon Restrepo, Jeremiah J. Zartman, and Konrad Basler

12.  Cultivation and Live Imaging of Drosophila Ovaries
Maureen Cetera, Lindsay Lewellyn, and Sally Horne-Badovinac

13.  Segmentation and Quantitative Analysis of Epithelial Tissues
Benoit Aigouy, Daiki Umetsu, and Suzanne Eaton

14.  Laser Ablation to Probe the Epithelial Mechanics in Drosophila
Pruthvi C. Shivakumar and Pierre-Francois Lenne

15.  Rapid Ovary Mass-isolation (ROMi) to Obtain Large Quantities of Drosophila Egg Chambers for Fluorescent In-Situ Hybridization
Helena Jambor, Pavel Mejstrik, and Pavel Tomancak

16.  Chromatin Immunoprecipitation for Analyzing Transcription Factor Binding and Histone Modifications in Drosophila
Yad Ghavi-Helm, Bingqing Zhao, and Eileen E.M. Furlong
17.  Protocols to Study Growth and Metabolism in Drosophila
Katrin Strassburger and Aurelio A. Teleman

18.  Protocols to Study Ageing in Drosophila
Matthew D. W. Piper and Linda Partridge

19.  Protocols to Study Behavior in Drosophila
Wendi S. Neckameyer and Parag Bhatt

20.  Electrophysiological Recordings from Lobula Plate Tangential Cells in Drosophila
Alex S. Mauss and Alexander Borst

21.  Methods to Establish a Drosophila Cell Line
Alain Debec, Timothy L. Megraw, and Antoine Guichet