Site-Specific Cassette Exchange Systems in the Aedes aegypti Mosquito and the Plutella xylostella Moth

Haghighat-Khah, Roya Elaine; Scaife, Sarah; Martins, Sara; John, Oliver St; Matzen, Kelly; Morrison, Neil I.; Alphey, Luke

doi:10.1371/journal.pone.0121097

Cited by 30 publications

(44 citation statements)

References 64 publications

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“…Considering the results presented here and the previous occurrence of single-site recombinants in Cre-, FLP, and phiC31-RMCE systems and the demonstration of their functionality as RMCE intermediates17, however, the formation of a stable intermediate reaction (i.e. integration) product might be a more common event in RMCE than previously assumed and points towards successive instead of simultaneous crossovers.…”

Section: Discussionsupporting

confidence: 46%

Cre/lox-Recombinase-Mediated Cassette Exchange for Reversible Site-Specific Genomic Targeting of the Disease Vector, Aedes aegypti

Häcker

Harrell

Eichner

et al. 2017

Sci Rep

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Site-specific genome modification (SSM) is an important tool for mosquito functional genomics and comparative gene expression studies, which contribute to a better understanding of mosquito biology and are thus a key to finding new strategies to eliminate vector-borne diseases. Moreover, it allows for the creation of advanced transgenic strains for vector control programs. SSM circumvents the drawbacks of transposon-mediated transgenesis, where random transgene integration into the host genome results in insertional mutagenesis and variable position effects. We applied the Cre/lox recombinase-mediated cassette exchange (RMCE) system to Aedes aegypti, the vector of dengue, chikungunya, and Zika viruses. In this context we created four target site lines for RMCE and evaluated their fitness costs. Cre-RMCE is functional in a two-step mechanism and with good efficiency in Ae. aegypti. The advantages of Cre-RMCE over existing site-specific modification systems for Ae. aegypti, phiC31-RMCE and CRISPR, originate in the preservation of the recombination sites, which 1) allows successive modifications and rapid expansion or adaptation of existing systems by repeated targeting of the same site; and 2) provides reversibility, thus allowing the excision of undesired sequences. Thereby, Cre-RMCE complements existing genomic modification tools, adding flexibility and versatility to vector genome targeting.

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Section: Discussionsupporting

confidence: 46%

Cre/lox-Recombinase-Mediated Cassette Exchange for Reversible Site-Specific Genomic Targeting of the Disease Vector, Aedes aegypti

Häcker

Harrell

Eichner

et al. 2017

Sci Rep

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“…The practical application of new technologies increases our understanding of the regulatory mechanisms underlying colour pattern divergence. Transgenic techniques have also been applied to construct transgenic strains in other lepidopteran species (Marcus et al ., ; Haghighat‐Khah et al ., ). Although previous studies have not investigated the regulatory region of pigmentation genes in lepidopteran insects, the regulatory regions of upstream regulatory genes that govern pattern formation, including transcription factors such as distal‐less (Carroll et al ., ; Brakefield et al ., ; Zhang and Reed, ), optix (Reed et al ., ), cortex (Nadeau et al ., ) ; and morphogens such as wingless (Martin and Reed, ; Yamaguchi et al ., ) and wntA (Martin et al ., ; Gallant et al ., ; Martin and Reed, ) were suggested.…”

Section: Discussionmentioning

confidence: 98%

Modular cis‐regulatory logic of yellow gene expression in silkmoth larvae

Suzuki

Koshikawa

Kobayashi

et al. 2019

Insect Molecular Biology

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Colour patterns in butterflies and moths are crucial traits for adaptation. Previous investigations have highlighted genes responsible for pigmentation (ie yellow and ebony). However, the mechanisms by which these genes are regulated in lepidopteran insects remain poorly understood. To elucidate this, molecular studies involving dipterans have largely analysed the cis‐regulatory regions of pigmentation genes and have revealed cis‐regulatory modularity. Here, we used well‐developed transgenic techniques in Bombyx mori and demonstrated that cis‐regulatory modularity controls tissue‐specific expression of the yellow gene. We first identified which body parts are regulated by the yellow gene via black pigmentation. We then isolated three discrete regulatory elements driving tissue‐specific gene expression in three regions of B. mori larvae. Finally, we found that there is no apparent sequence conservation of cis‐regulatory regions between B. mori and Drosophila melanogaster, and no expression driven by the regulatory regions of one species when introduced into the other species. Therefore, the trans‐regulatory landscapes of the yellow gene differ significantly between the two taxa. The results of this study confirm that lepidopteran species use cis‐regulatory modules to control gene expression related to pigmentation, and represent a powerful cadre of transgenic tools for studying evolutionary developmental mechanisms.

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“…Insertion of foreign genes into mosquito genomes was achieved by the use of Drosophila transposable P elements. Because of the limited capability of carrying large genetic elements and lack of site-specific integration, modified methods with bacteriophage Cre, phiC31, and yeast flippase (FLP) recombinases were further developed [138,139,140,141]. Other transposable elements isolated from other arthropod species remain as popular options for developing methods for transforming mosquitoes [142].…”

Section: Genetically Modified Mosquitoes For Vector Controlmentioning

confidence: 99%

Biological Control Strategies for Mosquito Vectors of Arboviruses

Huang

Higgs

Vanlandingham

2017

Insects

119

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Historically, biological control utilizes predatory species and pathogenic microorganisms to reduce the population of mosquitoes as disease vectors. This is particularly important for the control of mosquito-borne arboviruses, which normally do not have specific antiviral therapies available. Although development of resistance is likely, the advantages of biological control are that the resources used are typically biodegradable and ecologically friendly. Over the past decade, the advancement of molecular biology has enabled optimization by the manipulation of genetic materials associated with biological control agents. Two significant advancements are the discovery of cytoplasmic incompatibility induced by Wolbachia bacteria, which has enhanced replacement programs, and the introduction of dominant lethal genes into local mosquito populations through the release of genetically modified mosquitoes. As various arboviruses continue to be significant public health threats, biological control strategies have evolved to be more diverse and become critical tools to reduce the disease burden of arboviruses.

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Site-Specific Cassette Exchange Systems in the Aedes aegypti Mosquito and the Plutella xylostella Moth

Cited by 30 publications

References 64 publications

Cre/lox-Recombinase-Mediated Cassette Exchange for Reversible Site-Specific Genomic Targeting of the Disease Vector, Aedes aegypti

Cre/lox-Recombinase-Mediated Cassette Exchange for Reversible Site-Specific Genomic Targeting of the Disease Vector, Aedes aegypti

Modular cis‐regulatory logic of yellow gene expression in silkmoth larvae

Biological Control Strategies for Mosquito Vectors of Arboviruses

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