Main description:

This volume explores the latest techniques used to study Leishmania and other trypanosomatidae. Chapters in this book cover a wide range of topics such as cultivation of axenic amastigote forms; phylogeny and comparative genomics; and in vitro and in vivo infection models that look at hosts ranging from cell lines to mammalian and arthropode hosts. 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.

Unique and cutting-edge, Leishmania: Methods and Protocols is a valuable guide for both established and novice researchers in the Leishmania field. This book will also be useful to researchers working with other protozoa of the Order Trypanosomatide.

Contents:

Preface...Table of Contents...Contributing Authors...
1. Host-Free Systems for Differentiation of Axenic LeishmaniaDan Zilberstein and Roni Nitzan Koren
2. Phylogenetic StudiesKatrin Kuhls and Isabel Mauricio
3. A Guide to Next Generation Sequence Analysis of Leishmania GenomesHideo Imamura and Jean-Claude Dujardin
4. Quantitative RNA Analysis using RNA SeqPeter J. Myler, Jacqueline A. McDonald, Pedro Alcolea, and Aakash Sur
5. Ribosome Profiling in TrypanosomatidsAmelie J. Kraus and Raul O. Cosentino
6. Cosmid Library Construction and Functional CloningJoachim Clos and Dorotheo Zander-Dinse
7. Cos-Seq, A High Throughput Gain of Function Screen for Drug Resistance Studies in LeishmaniaJade-Eva Potvin, Philippe Leprohon, Elodie Gazanion, Mansi Sharma, Christopher Fernandez-Prada, and Marc Ouellette
8. Gene Replacement by Homologous RecombinationHenner Zirpel and Joachim Clos
9. LeishGEdit: A Method for Rapid Gene Knockout and Tagging using CRISPR-Cas9Tom Beneke and Eva Gluenz
10. DiCre-Based Inducible Disruption of Leishmania GenesSamuel M. Duncan, Elmarie Myburgh, Eliza Alves-Ferreira, and Jeremy C. Mottram
11. DiCre-Based Inducible Gene ExpressionJeziel D. Damasceno, Luiz R. O. Tosi, Renato E. R. S. Santos, and Jeremy C. Mottram
12. Generation of Bone Marrow-Derived Macrophages for In Vitro Infection ExperimentsEugenia Bifeld
13. Quantification of Intracellular Leishmania spp. Using Real-Time Quantitative PCR (qPCR)Eugenia Bifeld
14. In Vitro Infections of Macrophage-Like Cell Lines with Leishmania infantum for Drug ScreeningNuno Santarem, Joana Tavares, and Anabela Cordeiro-da-Silva
15. Quantification of Parasite Loads by Automated Microscopic Image AnalysisCarolina Borsoi Moraes and Laura Maria Alcantara
16. Quantification of Leishmania Parasites in Murine Models of Visceral InfectionJoana Tavares, Nuno Santarem, and Anabela Cordeiro-da-Silva
17. Syrian Hamster as an Advanced Experimental Model for Visceral LeishmaniasisMaria Dolores Jimenez-Anton, Montserrat Grau, Ana Isabel Olias-Molero, and Jose M Alunda
18. Experimental Cutaneous Leishmaniasis: Mouse Models for Resolution of Inflammation versus Chronicity of DiseaseChristian Bogdan, Andrea Debus, Heidi Sebald, Bapli Rai, Johanna Schafer, Stephanie Obermeyer, and Ulrike Schleicher
19. Establishment, Maintenance of Phlemotomus spp. In the Laboratory, and Infection with Leishmania spp.Ifhem Chelbi and Elyes Zhioua