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2 results on '"MESH : Anopheles"'

1. Efficient PhiC31 integrase-mediated site-specific germline transformation of Anopheles gambiae

Author

Kenneth D. Vernick, Nicolas Puchot, Catherine Bourgouin, Gareth J Lycett, Emilie Pondeville, Amy Lynd, J. M. Meredith, Paul Eggleston, Génétique et Génomique des Insectes vecteurs, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Keele University [Keele], Liverpool School of Tropical Medicine (LSTM), Support to E.P. was from an ANR-07-MIME-O25-01 award to C.B., from the Roux Foundation (Pasteur Institute) and from a UE323173 Anopath award to K.D.V., to C.B. from award no. ANR-10-LABX-62-IBEID, to P.E. from a Wellcome Trust Programme grant (0084582) and the EU FP7 (INFRAVEC), and to G.J.L. from European Commission grant no. 223736 (TransMalariaBloc) and a Biotechnology and Biological Sciences Research Council grant (no. BB/F021933/1)., We are grateful to the Photography Department of Institut Pasteur, especially F. Gardy and J.-M. Panaud for the production of the video for embryo alignment, and to M. Calos for pET11-ΦC31 and original attP and attB constructs. We thank A. James, A. Handler and N. Windbichler for helpful technical discussions., ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), European Project: 323173,EC:FP7:ERC,ERC-2012-ADG_20120314,ANOPATH(2013), European Project: 228421,EC:FP7:INFRA,FP7-INFRASTRUCTURES-2008-1,INFRAVEC(2009), European Project: 223736,EC:FP7:HEALTH,FP7-HEALTH-2007-B,TRANSMALARIABLOC(2008), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Liverpool School of Tropical Medicine ( LSTM ), ANR-10-LABX-62-IBEID,IBEID,Laboratoire d'Excellence 'Integrative Biology of Emerging Infectious Diseases' ( 2010 ), European Project : 323173,EC:FP7:ERC,ERC-2012-ADG_20120314,ANOPATH ( 2013 ), European Project : 228421,EC:FP7:INFRA,FP7-INFRASTRUCTURES-2008-1,INFRAVEC ( 2009 ), European Project : 223736,EC:FP7:HEALTH,FP7-HEALTH-2007-B,TRANSMALARIABLOC ( 2008 ), and Centre National de la Recherche Scientifique ( CNRS ) -Institut Pasteur [Paris]

Subjects
MESH : RNA, Messenger, MESH: Integrases, [ SDV.BA.ZI ] Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, Anopheles gambiae, MESH : Animals, Genetically Modified, Messenger/metabolism, Genome, Germline, Animals, Genetically Modified, Genetic Engineering/*methods, Plasmid, MESH: Animals, Genetics, [ SDV.SPEE ] Life Sciences [q-bio]/Santé publique et épidémiologie, 3. Good health, Genetically modified organism, Transformation, MESH : Anopheles, Larva, MESH: Genetic Engineering, Genetic Engineering, Functional genomics, [ SDV.MP.PAR ] Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology, Transgene, Genetically Modified, MESH: Transformation, Genetic, Biology, Integrases/*genetics, General Biochemistry, Genetics and Molecular Biology, MESH : Genetic Engineering, MESH: Animals, Genetically Modified, MESH: Anopheles, MESH : Integrases, Transformation, Genetic, Genetic, Anopheles, parasitic diseases, Animals, [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology, RNA, Messenger, Anopheles/*genetics, MESH: RNA, Messenger, MESH : Larva, Integrases, fungi, Larva/genetics, biology.organism_classification, [SDV.BA.ZI]Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, Transformation (genetics), RNA, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, MESH : Animals, MESH : Transformation, Genetic, MESH: Larva
Abstract

International audience; Current transgenic methodology developed for mosquitoes has not been applied widely to the major malaria vector Anopheles gambiae, which has proved more difficult to genetically manipulate than other mosquito species and dipteran insects. In this protocol, we describe PhiC31-mediated site-specific integration of transgenes into the genome of A. gambiae. The PhiC31 system has many advantages over 'classical' transposon-mediated germline transformation systems, because it allows integration of large transgenes at specific, characterized genomic locations. Starting from a general protocol, we have optimized steps from embryo collection to co-injection of transgene-containing plasmid and in vitro-produced PhiC31 integrase mRNA. We also provide tips for screening transgenic larvae. The outlined procedure provides robust transformation in A. gambiae, resulting in homozygous transgenic lines in approximately 2-3 months.

Published
2014
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2. IgG responses to the gSG6-P1 salivary peptide for evaluating human exposure to Anopheles bites in urban areas of Dakar region, Sénégal

Author

Christophe Rogier, Sylvie Cornelie, Papa M. Drame, Stéphanie Dos Santos, Frédéric Pagès, Mbacké Sembène, Vanessa Machault, A. Diallo, Richard Lalou, Anne Poinsignon, Franck Remoue, Jean-Yves Le Hesran, Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle ( MIVEGEC ), Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD [France-Sud] ), Laboratoire d'aérologie - LA ( LA ), Université Toulouse III - Paul Sabatier ( UPS ), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut national des sciences de l'Univers ( INSU - CNRS ) -Observatoire Midi-Pyrénées ( OMP ) -Centre National de la Recherche Scientifique ( CNRS ), Centre National d'Etudes Spatiales ( CNES ), Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes ( URMITE ), Institut de Recherche pour le Développement ( IRD ) -Aix Marseille Université ( AMU ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -IFR48, INSB-INSB-Centre National de la Recherche Scientifique ( CNRS ), Santé de la mère et de l'enfant en milieu tropical : épidémiologie génétique et périnatale, Université Paris Descartes - Paris 5 ( UPD5 ), Laboratoire Population-Environnement-Développement ( LPED ), Institut de Recherche pour le Développement ( IRD ) -Aix Marseille Université ( AMU ), Faculté des Sciences et Techniques (FST), Université Cheikh Anta Diop ( UCAD ), Institut de recherche pour le développement ( IRD [Sénégal] ), Institut Pasteur de Madagascar, Institut Pasteur de Madagascar-Réseau International des Instituts Pasteur ( RIIP ), PMD was supported by a PhD scholarship provided by IRD (DSF Department) then a fellowship from InfectioPole Sud Fundation. VM received financial support from the Direction Générale de l'Armement (DGA-Contrat d'Objectif n°07CO402) and the Centre National d'Etudes Spatiales (CNES). AD is a PhD student supported by IRDDSF Department. AP was supported by a fellowship from F. Lacoste., Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle (MIVEGEC), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud]), Laboratoire d'aérologie (LAERO), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Observatoire Midi-Pyrénées (OMP), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Météo-France -Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Météo-France -Centre National de la Recherche Scientifique (CNRS), Centre National d'Études Spatiales [Toulouse] (CNES), Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes (URMITE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR48, Institut des sciences biologiques (INSB-CNRS)-Institut des sciences biologiques (INSB-CNRS)-Centre National de la Recherche Scientifique (CNRS), Université Paris Descartes - Paris 5 (UPD5), Vector Control Group (MIVEGEC-VCG), Evolution des Systèmes Vectoriels (ESV), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud])-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud])-Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle (MIVEGEC), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud])-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud]), Transmission-Interactions-Adaptations hôtes/vecteurs/pathogènes (MIVEGEC-TRIAD), Laboratoire Population-Environnement-Développement (LPED), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU), Université Cheikh Anta Diop [Dakar, Sénégal] (UCAD), Institut de recherche pour le développement (IRD [Sénégal]), Réseau International des Instituts Pasteur (RIIP), Institut de Recherche pour le Développement (IRD [France-Sud])-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Laboratoire d'aérologie (LA), Centre National de la Recherche Scientifique (CNRS)-Observatoire Midi-Pyrénées (OMP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, INSB-INSB-Centre National de la Recherche Scientifique (CNRS), and Météo France-Centre National d'Études Spatiales [Toulouse] (CNES)-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Météo France-Centre National d'Études Spatiales [Toulouse] (CNES)-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3)

Subjects
Male, Urban Population, Anopheles gambiae, [SDV]Life Sciences [q-bio], MESH : Analysis of Variance, MESH : Child, Preschool, 0302 clinical medicine, MESH : Child, MESH: Child, MESH : Insect Vectors, MESH: Animals, MESH : Female, 030212 general & internal medicine, Child, 2. Zero hunger, MESH: Immunoglobulin G, education.field_of_study, biology, MESH : Salivary Proteins and Peptides, Transmission (medicine), Anopheles, MESH: Salivary Proteins and Peptides, Environmental exposure, MESH : Adult, MESH : Senegal, Senegal, MESH: Antibody Formation, 3. Good health, MESH: Urban Population, Infectious Diseases, MESH : Anopheles, MESH: Young Adult, Child, Preschool, MESH : Urban Population, Female, Adult, lcsh:Arctic medicine. Tropical medicine, MESH : Immunoglobulin G, lcsh:RC955-962, MESH : Male, 030231 tropical medicine, Population, MESH: Environmental Exposure, MESH : Young Adult, MESH: Insect Vectors, MESH : Antibody Formation, Insect bites and stings, lcsh:Infectious and parasitic diseases, MESH: Anopheles, Young Adult, 03 medical and health sciences, MESH: Insect Bites and Stings, MESH: Senegal, MESH : Insect Bites and Stings, Environmental health, MESH: Analysis of Variance, parasitic diseases, medicine, Animals, Humans, lcsh:RC109-216, Salivary Proteins and Peptides, education, Analysis of Variance, MESH: Humans, [ SDV ] Life Sciences [q-bio], Research, MESH: Child, Preschool, MESH : Humans, Insect Bites and Stings, MESH: Adult, Environmental Exposure, biology.organism_classification, medicine.disease, MESH: Male, Insect Vectors, Parasitology, Immunoglobulin G, Antibody Formation, Immunology, MESH : Animals, MESH: Female, MESH : Environmental Exposure, Malaria
Abstract

Background Urban malaria can be a serious public health problem in Africa. Human-landing catches of mosquitoes, a standard entomological method to assess human exposure to malaria vector bites, can lack sensitivity in areas where exposure is low. A simple and highly sensitive tool could be a complementary indicator for evaluating malaria exposure in such epidemiological contexts. The human antibody response to the specific Anopheles gSG6-P1 salivary peptide have been described as an adequate tool biomarker for a reliable assessment of human exposure level to Anopheles bites. The aim of this study was to use this biomarker to evaluate the human exposure to Anopheles mosquito bites in urban settings of Dakar (Senegal), one of the largest cities in West Africa, where Anopheles biting rates and malaria transmission are supposed to be low. Methods One cross-sectional study concerning 1,010 (505 households) children (n = 505) and adults (n = 505) living in 16 districts of downtown Dakar and its suburbs was performed from October to December 2008. The IgG responses to gSG6-P1 peptide have been assessed and compared to entomological data obtained in or near the same district. Results Considerable individual variations in anti-gSG6-P1 IgG levels were observed between and within districts. In spite of this individual heterogeneity, the median level of specific IgG and the percentage of immune responders differed significantly between districts. A positive and significant association was observed between the exposure levels to Anopheles gambiae bites, estimated by classical entomological methods, and the median IgG levels or the percentage of immune responders measuring the contact between human populations and Anopheles mosquitoes. Interestingly, immunological parameters seemed to better discriminate the exposure level to Anopheles bites between different exposure groups of districts. Conclusions Specific human IgG responses to gSG6-P1 peptide biomarker represent, at the population and individual levels, a credible new alternative tool to assess accurately the heterogeneity of exposure level to Anopheles bites and malaria risk in low urban transmission areas. The development of such biomarker tool would be particularly relevant for mapping and monitoring malaria risk and for measuring the efficiency of vector control strategies in these specific settings.

Published
2012
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