Journal ArticleDOI
Germline transformation of the malaria vector, Anopheles gambiae, with the piggyBac transposable element.
Genelle L. Grossman,Cristina S. Rafferty,John R. Clayton,T. K. Stevens,O. Mukabayire,Mark Q. Benedict +5 more
TLDR
An improvement in the injection technique at the end of the studies resulted in increased G0 hatching, transient expression and EGFP‐expression rates among G1 progeny, and genetic cross data suggest that the insertion site of the element either resulted in, or is tightly linked to, a recessive lethal.Abstract:
Germline transformation of the major African malaria vector, Anopheles gambiae, was achieved using the piggyBac transposable element marked with the enhanced green fluorescent protein (EGFP) injected into mosquito embryos. Two G1 generation male mosquitoes expressing EGFP were identified among 34 143 larvae screened. Genomic Southern data and sequencing of the piggyBac insertion boundaries showed that these two males arose from one piggyBac insertion event in the injected G0 embryos. Genetic cross data suggest that the insertion site of the element either resulted in, or is tightly linked to, a recessive lethal. This was demonstrated by a deficiency in the number of EGFP-expressing offspring from inbred crosses but expected ratios in outcrosses to non-transformed individuals and failure to establish a pure-breeding line. The insertion was weakly linked to the collarless locus on chromosome 2 and was shown by in situ hybridization to be located in division 28D of that chromosome. Particularly high levels of expression were observed uniformly in salivary glands and, in most individuals, in the anterior stomach. An improvement in the injection technique at the end of the studies resulted in increased G0 hatching, transient expression and EGFP-expression rates among G1 progeny.read more
Citations
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Journal ArticleDOI
Immunity-Related Genes and Gene Families in Anopheles gambiae
George K. Christophides,Evgeny M. Zdobnov,Carolina Barillas-Mury,Ewan Birney,Stéphanie Blandin,Claudia Blass,Paul T. Brey,Frank H. Collins,Alberto Danielli,George Dimopoulos,Charles Hetru,Ngo Thi Hoa,Jules A. Hoffmann,Stefan M. Kanzok,Ivica Letunic,Elena A. Levashina,Thanasis G. Loukeris,Gareth J Lycett,Stephan Meister,Kristin Michel,Luis F. Moita,Hans-Michael Müller,Mike A. Osta,Susan M. Paskewitz,Jean-Marc Reichhart,Andrey Rzhetsky,Laurent Troxler,Kenneth D. Vernick,Dina Vlachou,Jennifer Volz,Christian von Mering,Jiannong Xu,Liangbiao Zheng,Peer Bork,Fotis C. Kafatos +34 more
TL;DR: 242 Anopheles gambiae genes from 18 gene families implicated in innate immunity are identified and marked diversification relative to Drosophila melanogaster is detected, confirming that sequence diversification is accompanied by specific responses to different immune challenges.
Journal ArticleDOI
Gene drive systems for insect disease vectors
Steven P. Sinkins,Fred Gould +1 more
TL;DR: The potential of natural gene drive systems for spreading genes that can block the transmission of insect-borne pathogens are explored and the artificial constructs that could be envisaged for this purpose are discussed.
Journal ArticleDOI
Bacteria of the genus Asaia stably associate with Anopheles stephensi, an Asian malarial mosquito vector
Guido Favia,Irene Ricci,Claudia Damiani,Noura Raddadi,Elena Crotti,Massimo Marzorati,Aurora Rizzi,Roberta Urso,Lorenzo Brusetti,Sara Borin,Diego Mora,Patrizia Scuppa,Luciano Pasqualini,E. Clementi,Marco Genchi,S. Corona,Ilaria Negri,Giulio Grandi,Alberto Alma,Laura D. Kramer,Fulvio Esposito,Claudio Bandi,Luciano Sacchi,Daniele Daffonchio +23 more
TL;DR: It is shown that an α-proteobacterium of the genus Asaia is stably associated with larvae and adults of Anopheles stephensi, an important mosquito vector of Plasmodium vivax, a main malaria agent in Asia.
Journal ArticleDOI
Reverse genetics in the mosquito Anopheles gambiae: targeted disruption of the Defensin gene
Stéphanie Blandin,Luis F. Moita,Thomas Köcher,Matthias Wilm,Fotis C. Kafatos,Elena A. Levashina +5 more
TL;DR: A simple in vivo double‐stranded RNA knockout approach to determine the function of the mosquito antimicrobial peptide gene Defensin concludes that this peptide is not a major antiparasitic factor in A. gambiae in vivo.
Journal ArticleDOI
Malaria Control with Genetically Manipulated Insect Vectors
Luke Alphey,C. Ben Beard,Peter F. Billingsley,Maureen Coetzee,Andrea Crisanti,C. F. Curtis,Paul Eggleston,Charles Godfray,Janet Hemingway,Marcelo Jacobs-Lorena,Anthony A. James,Fotis C. Kafatos,Louis G. Mukwaya,Michael Paton,Jeffrey R. Powell,William Schneider,Thomas W. Scott,Barbara Sina,Robert E. Sinden,Steven P. Sinkins,Andrew Spielman,Yeya T. Touré,Frank H. Collins +22 more
TL;DR: At a recent workshop, experts discussed the benefits, risks, and research priorities associated with using genetically manipulated insects in the control of vector-borne diseases.
References
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Journal ArticleDOI
Germline transformation of the silkworm Bombyx mori L. using a piggyBac transposon-derived vector
Toshiki Tamura,Chantal Thibert,Corinne Royer,Toshio Kanda,Abraham Eappen,Mari Kamba,Natuo Kômoto,Jean-Luc Thomas,Bernard Mauchamp,Gérard Chavancy,Paul D. Shirk,Malcolm J. Fraser,Jean-Claude Prudhomme,Pierre Couble +13 more
TL;DR: The presence of the inverted terminal repeats of piggyBac and the characteristic TTAA sequence at the borders of all the analyzed inserts confirmed that transformation resulted from precise transposition events.
Journal ArticleDOI
A ribosomal RNA gene probe differentiates member species of the Anopheles gambiae complex.
TL;DR: A 0.59 kilobase DNA fragment cloned from an rDNA cistron of the mosquito Anopheles gambiae can be used as a probe to differentiate between A. gambiae, A. arabiensis, and A. melas, three morphologically identical sibling species in the A. gamblingiae complex which otherwise can be reliably distinguished only by polytene chromosome banding patterns.
Journal ArticleDOI
Insect population control using a dominant, repressible, lethal genetic system.
TL;DR: A major modification to the sterile insect technique is described, in which transgenic insects homozygous for a dominant, repressible, female-specific lethal gene system are used.
Journal ArticleDOI
Transposon mutagenesis of baculoviruses: analysis of Trichoplusia ni transposon IFP2 insertions within the FP-locus of nuclear polyhedrosis viruses.
Lynne Csiszar Cary,Michael Goebel,Bartholomew G. Corsaro,Hwei Gene Wang,Elliot D. Rosen,Malcolm J. Fraser +5 more
TL;DR: The transposable IFP2 element of Trichoplusia ni was originally isolated as a host DNA insertion in spontaneous FP mutants of Galleria mellonella or Autographa californica nuclear polyhedrosis viruses, but is not apparent in DNAs isolated from the TN-R2 cell line or the authors' laboratory colony of T. ni larvae, suggesting IFP1 was recently introduced into the T. Ni genome.
Journal ArticleDOI
High-frequency P element loss in Drosophila is homolog dependent.
TL;DR: A model of P element transposition is suggested in which jumps occur by a "cut-and-paste" mechanism but are followed by double-strand gap repair to restore the P element at the donor site.
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