Optimising homing endonuclease gene drive performance in a semi-refractory species: the Drosophila melanogaster experience.

Optimising homing endonuclease gene drive performance in a semi-refractory species: the Drosophila melanogaster experience.

Homing endonuclease gene (HEG) drive is a promising insect population control technique that employs meganucleases to impair the fitness of pest populations. Our previous studies showed that HEG drive was more difficult to achieve in Drosophila melanogaster than Anopheles gambiae and we therefore in...

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Autores principales: Yuk-Sang Chan, David S Huen, Ruth Glauert, Eleanor Whiteway, Steven Russell
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Publicado: Public Library of Science (PLoS) 2013
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Acceso en línea:https://doaj.org/article/cbd2b010179a4767b847a69147febc5c
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spelling oai:doaj.org-article:cbd2b010179a4767b847a69147febc5c2021-11-18T08:00:53ZOptimising homing endonuclease gene drive performance in a semi-refractory species: the Drosophila melanogaster experience.1932-620310.1371/journal.pone.0054130https://doaj.org/article/cbd2b010179a4767b847a69147febc5c2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23349805/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Homing endonuclease gene (HEG) drive is a promising insect population control technique that employs meganucleases to impair the fitness of pest populations. Our previous studies showed that HEG drive was more difficult to achieve in Drosophila melanogaster than Anopheles gambiae and we therefore investigated ways of improving homing performance in Drosophila. We show that homing in Drosophila responds to increased expression of HEGs specifically during the spermatogonia stage and this could be achieved through improved construct design. We found that 3'-UTR choice was important to maximise expression levels, with HEG activity increasing as we employed Hsp70, SV40, vasa and βTub56D derived UTRs. We also searched for spermatogonium-specific promoters and found that the Rcd-1r promoter was able to drive specific expression at this stage. Since Rcd-1 is a regulator of differentiation in other species, it suggests that Rcd-1r may serve a similar role during spermatogonial differentiation in Drosophila. Contrary to expectations, a fragment containing the entire region between the TBPH gene and the bgcn translational start drove strong HEG expression only during late spermatogenesis rather than in the germline stem cells and spermatogonia as expected. We also observed that the fraction of targets undergoing homing was temperature-sensitive, falling nearly four-fold when the temperature was lowered to 18°C. Taken together, this study demonstrates how a few simple measures can lead to substantial improvements in the HEG-based gene drive strategy and reinforce the idea that the HEG approach may be widely applicable to a variety of insect control programs.Yuk-Sang ChanDavid S HuenRuth GlauertEleanor WhitewaySteven RussellPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 1, p e54130 (2013)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Yuk-Sang Chan
David S Huen
Ruth Glauert
Eleanor Whiteway
Steven Russell
Optimising homing endonuclease gene drive performance in a semi-refractory species: the Drosophila melanogaster experience.
description Homing endonuclease gene (HEG) drive is a promising insect population control technique that employs meganucleases to impair the fitness of pest populations. Our previous studies showed that HEG drive was more difficult to achieve in Drosophila melanogaster than Anopheles gambiae and we therefore investigated ways of improving homing performance in Drosophila. We show that homing in Drosophila responds to increased expression of HEGs specifically during the spermatogonia stage and this could be achieved through improved construct design. We found that 3'-UTR choice was important to maximise expression levels, with HEG activity increasing as we employed Hsp70, SV40, vasa and βTub56D derived UTRs. We also searched for spermatogonium-specific promoters and found that the Rcd-1r promoter was able to drive specific expression at this stage. Since Rcd-1 is a regulator of differentiation in other species, it suggests that Rcd-1r may serve a similar role during spermatogonial differentiation in Drosophila. Contrary to expectations, a fragment containing the entire region between the TBPH gene and the bgcn translational start drove strong HEG expression only during late spermatogenesis rather than in the germline stem cells and spermatogonia as expected. We also observed that the fraction of targets undergoing homing was temperature-sensitive, falling nearly four-fold when the temperature was lowered to 18°C. Taken together, this study demonstrates how a few simple measures can lead to substantial improvements in the HEG-based gene drive strategy and reinforce the idea that the HEG approach may be widely applicable to a variety of insect control programs.
format article
author Yuk-Sang Chan
David S Huen
Ruth Glauert
Eleanor Whiteway
Steven Russell
author_facet Yuk-Sang Chan
David S Huen
Ruth Glauert
Eleanor Whiteway
Steven Russell
author_sort Yuk-Sang Chan
title Optimising homing endonuclease gene drive performance in a semi-refractory species: the Drosophila melanogaster experience.
title_short Optimising homing endonuclease gene drive performance in a semi-refractory species: the Drosophila melanogaster experience.
title_full Optimising homing endonuclease gene drive performance in a semi-refractory species: the Drosophila melanogaster experience.
title_fullStr Optimising homing endonuclease gene drive performance in a semi-refractory species: the Drosophila melanogaster experience.
title_full_unstemmed Optimising homing endonuclease gene drive performance in a semi-refractory species: the Drosophila melanogaster experience.
title_sort optimising homing endonuclease gene drive performance in a semi-refractory species: the drosophila melanogaster experience.
publisher Public Library of Science (PLoS)
publishDate 2013
url https://doaj.org/article/cbd2b010179a4767b847a69147febc5c
work_keys_str_mv AT yuksangchan optimisinghomingendonucleasegenedriveperformanceinasemirefractoryspeciesthedrosophilamelanogasterexperience
AT davidshuen optimisinghomingendonucleasegenedriveperformanceinasemirefractoryspeciesthedrosophilamelanogasterexperience
AT ruthglauert optimisinghomingendonucleasegenedriveperformanceinasemirefractoryspeciesthedrosophilamelanogasterexperience
AT eleanorwhiteway optimisinghomingendonucleasegenedriveperformanceinasemirefractoryspeciesthedrosophilamelanogasterexperience
AT stevenrussell optimisinghomingendonucleasegenedriveperformanceinasemirefractoryspeciesthedrosophilamelanogasterexperience
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