Genetic sexing through the use of Y-linked transgenes

Condon, Kirsty C; Condon, George C.; Dafa'alla, Tarig; Fu, Guoliang; Phillips, Caroline E.; Jin, Li; Gong, Peng; Alphey, Luke

doi:10.1016/j.ibmb.2007.07.006

Cited by 36 publications

(24 citation statements)

References 44 publications

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“…Nevertheless, Y-linked fluorescence does provide the possibility for male-specific selection, especially if embryonic expression occurs (or is detectable) for fluorescence-based embryonic sorting. A Y-linked insertion of a fluorescent protein-marked vector has been reported for the medfly, Ceratitis capitata (Condon et al 2007a) and here we report the first Y-linked vector insertion in A. ludens.…”

Section: Discussionmentioning

confidence: 54%

Development of transgenic strains for the biological control of the Mexican fruit fly, Anastrepha ludens

et al. 2010

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The Mexican fruit fly, Anastrepha ludens, is a highly significant agricultural pest species that has been genetically transformed with a piggyBac-based transposon vector system using independent vector and transposase helper plasmids. Minimum estimated germ-line transformation frequencies were approximately 13-21% per fertile G(0) individual, similar to previously reported frequencies using single vector-helper plasmids. Two vector constructs were tested with potential importance to transgenic strain development for mexfly biological control. The first allows post-integration stabilization of a transposon-vector by deletion of a terminal sequence necessary for mobilization. The complete pB[L1-EGFP-L2-DsRed-R1] vector was integrated into the Chiapas wild type strain with subsequent deletion of the L2-DsRed-R1 sub-vector carrying the piggyBac 3' terminal sequence. Quality control tests for three of the stabilization vector lines (previous to stabilization) assessed viability at all life stages, fertility, adult flight ability, and adult male sexual competitiveness. All three transgenic lines were less fit compared to the wild strain by approximately 5-10% in most tests, however, there was no significant difference in sexual competitiveness which is the major prerequisite for optimal strain release. The second vector, pB[XL-EGFP, Asß2-tub-DsRed.T3], has the DsRed.T3 fluorescent protein reporter gene regulated by the A. suspensa Asß2-tubulin promoter, that resulted in testis and sperm-specific DsRed fluorescence in transgenic male mexflies. Fluorescent sperm bundles were unambiguously observed in the spermathecae of non-transgenic females mated to transgenic males. One transgenic line apparently had a male-specific Y-chromosome insertion, having potential use for sexing by fluorescent-embryo sorting. All transgenic lines expressed easily detectable and stable fluorescence in adults allowing their identification after trapping in the field.

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Section: Discussionmentioning

confidence: 54%

Development of transgenic strains for the biological control of the Mexican fruit fly, Anastrepha ludens

et al. 2010

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“…The marking of males by fluorescent proteins had been previously achieved in several other insects [20,22,27], but fluorescence did not appear before late larval stages, with the exception of one system for the Mediterranean fruit fly [21]. Because feeding larvae increases production costs in large-scale rearing, developing embryonic sexing systems remained a high priority.…”

Section: Discussionmentioning

confidence: 99%

“…Such strains have been developed in mosquitoes and fruit flies by inserting a transgene carrying a fluorescent protein under the control of a testis-specific or constitutive promoters [20][21][22][23]. Automated fluorescence sorters (COPAS, Union Biometrica) could then be used to separate male and female larvae.…”

Section: Introductionmentioning

confidence: 99%

Y-Linked Markers for Improved Population Control of the Tephritid Fruit Fly Pest, Anastrepha suspensa

Schetelig

Handler

2013

Yellow Biotechnology II

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Insect pest control programs incorporating the sterile insect technique (SIT) rely on the mass production and release of sterilized insects to reduce the wild-type population through infertile matings. Most effective programs release only males to avoid any crop damage caused by female fruit flies or transmission of disease by female mosquitoes. Therefore, the females have to be eliminated, preferably in an early developmental stage, during mass rearing. Different systems and techniques have been created for the sex separation of a few insect species. One of these is the transgenic sex-specific fluorescent protein marking of the insects with automated fluorescent-based sorting of the individuals to achieve sex separation. Here we describe the Y-linked integration of fluorescent markers driven by the widely active Drosophila melanogaster polyubiquitin promoter in the Caribfly, Anastrepha suspensa. Four strains with Y-linked integrations were established with one line expressing the DsRed fluorescent protein marker during embryogenesis. This line now has the possibility for use with automated sex separation in rearing, and the same transgene markers could be used in other insects for similar applications.

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“…Another proposed use of recombinant DNA methods is to engineer sex-specific expression of a visible marker, such as a fluorescent protein, allowing fluorescence-based sorting. 23,24 This method may be limited by the speed of available fluorescencebased sorters.…”

Section: 16mentioning

confidence: 99%

Interspecific Cross-Mating BetweenAedes aegyptiandAedes albopictusLaboratory Strains: Implication of Population Density on Mating Behaviors

Marcela

Hassan

Ahmad

et al. 2015

Journal of the American Mosquito Control Association

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Genetic sexing through the use of Y-linked transgenes

Cited by 36 publications

References 44 publications

Development of transgenic strains for the biological control of the Mexican fruit fly, Anastrepha ludens

Development of transgenic strains for the biological control of the Mexican fruit fly, Anastrepha ludens

Y-Linked Markers for Improved Population Control of the Tephritid Fruit Fly Pest, Anastrepha suspensa

Interspecific Cross-Mating BetweenAedes aegyptiandAedes albopictusLaboratory Strains: Implication of Population Density on Mating Behaviors

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