Liverpool School of Tropical Medicine View Institution's Website 7 articles published in JoVE Genetics Small-Cage Laboratory Trials of Genetically-Engineered Anopheline Mosquitoes Rebeca Carballar-Lejarazú1, Thai Binh Pham2, Vanessa Bottino-Rojas1, Adriana Adolfi1,3, Anthony A. James1,2 1Department of Microbiology & Molecular Genetics, University of California, Irvine, 2Department of Molecular Biology & Biochemistry, University of California, Irvine, 3Vector Biology Department, Liverpool School of Tropical Medicine The protocols reported here illustrate three alternative ways to assess the performance of genetically-engineered mosquitoes destined for vector control in laboratory-contained small cage trials. Each protocol is tailored to the specific modification the mosquito strain bears (gene drive or non-gene drive) and the types of parameters measured. Genetics Using the GAL4-UAS System for Functional Genetics in Anopheles gambiae Beth Crawford Poulton1, Fraser Colman1, Amalia Anthousi1,2,3, Linda Grigoraki1, Adriana Adolfi1, Amy Lynd1, Gareth John Lycett1 1Department of Vector Biology, Liverpool School of Tropical Medicine, 2IMBB FORTH, 3Department of Biology, University of Crete The bipartite GAL4-UAS system is a versatile tool for modification of gene expression in a controlled spatiotemporal manner which permits functional genetic analysis in Anopheles gambiae. The procedures described for using this system are a semi-standardized cloning strategy, sexing and screening of pupae for fluorescent protein markers and embryo fixation. Genetics Site-Directed φC31-Mediated Integration and Cassette Exchange in Anopheles Vectors of Malaria Adriana Adolfi1,2, Amy Lynd1, Gareth J. Lycett1, Anthony A. James2,3 1Vector Biology Department, Liverpool School of Tropical Medicine, 2Department of Microbiology & Molecular Genetics, University of California, 3Department of Molecular Biology & Biochemistry, University of California The protocol describes how to achieve site-directed modifications in the genome of Anopheles malaria mosquitoes using the φC31 system. Modifications described include both the integration and the exchange of transgenic cassettes in the genome of attP-bearing docking lines. Biology IR-TEx: An Open Source Data Integration Tool for Big Data Transcriptomics Designed for the Malaria Vector Anopheles gambiae Victoria A. Ingham1, Andrew Bennett2, Duo Peng3, Simon C. Wagstaff2, Hilary Ranson1 1Vector Biology, Liverpool School of Tropical Medicine, 2Research Computing Unit, Liverpool School of Tropical Medicine, 3Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health IR-TEx explores insecticide resistance-related transcriptional profiles in the species Anopheles gambiae. Provided here are full instructions for using the application, modifications for exploring multiple transcriptomic datasets, and using the framework to build an interactive database for collections of transcriptomic data from any organism, generated in any platform. Medicine Bronchoalveolar Lavage (BAL) for Research; Obtaining Adequate Sample Yield Andrea M. Collins1,2, Jamie Rylance3, Daniel G. Wootton4, Angela D. Wright3,5, Adam K. A. Wright1,3, Duncan G. Fullerton3,6, Stephen B. Gordon3 1Biomedical Research Centre in Microbial Diseases, National Institute for Health Research, 2Respiratory Infection Group, Royal Liverpool and Broadgreen University Hospital Trust, 3Respiratory Infection Group, Liverpool School of Tropical Medicine, 4Institute of Infection and Global Health, University of Liverpool, 5Comprehensive Local Research Network, Royal Liverpool and Broadgreen University Hospital Trust, 6Department of Respiratory Research, University Hospital Aintree We describe a research technique for fiberoptic bronchoscopy and bronchoalveolar lavage using low pressure suction. The technique is used to harvest immune cells from the lung bronchoalveolar surfaces. Local anesthetic and mild conscious sedation (midazolam) is used. Subjects tolerate the procedure well and experience minimal side effects. Immunology and Infection A Simple Protocol for Platelet-mediated Clumping of Plasmodium falciparum-infected Erythrocytes in a Resource Poor Setting Dumizulu L. Tembo1, Jacqui Montgomery1, Alister G. Craig2, Samuel C. Wassmer3 1Malawi-Liverpool-Wellcome Trust Clinical Research Programme, 2Liverpool School of Tropical Medicine, 3Department of Microbiology, Division of Medical Parasitology, New York University School of Medicine This method investigates the platelet-mediated clumping phenotype of Plasmodium falciparum-infected erythrocytes (pRBC) in clinical isolates. This is performed by isolating and co-incubating platelet-rich plasma and a suspension of pRBC. Medicine Experimental Human Pneumococcal Carriage Jenna F. Gritzfeld1, Angie D. Wright1,2,3, Andrea M. Collins1,2,4, Shaun H. Pennington1, Adam K.A. Wright5, Aras Kadioglu6, Daniela M. Ferreira1, Stephen B. Gordon1 1Respiratory Infection Group, Liverpool School of Tropical Medicine, 2Royal Liverpool and Broadgreen, University Hospital Trust, 3Comprehensive Local Research Network, 4NIHR Biomedical Research Centre in Microbial Diseases, Royal Liverpool and Broadgreen University Hospitals NHS Trust, 5Institute of Lung Health, Respiratory Biomedical Unit, University Hospitals of Leicester NHS Trust & University of Leicester, 6Department of Clinical Infection Microbiology & Immunology, Institute of Infection & Global Health, University of Liverpool Experimental human pneumococcal carriage offers a natural model of carriage and a potential model for use in vaccine development. This technique is valuable yet complex and involves clinical risk by introducing a pathogen into a human. We have developed a detailed protocol.