Robyn Raban - Academia.edu (original) (raw)

Papers by Robyn Raban

Scientific Reports, Jan 17, 2024

The resident human skin microbiome is responsible for the production of most of the human scents ... more The resident human skin microbiome is responsible for the production of most of the human scents that are attractive to mosquitoes. Hence, engineering the human skin microbiome to synthesize less of mosquito attractants or produce repellents could potentially reduce bites and prevent the transmission of deadly mosquito-borne pathogens. In order to further characterize the human skin volatilome, we quantified the major volatiles of 39 strains of skin commensals (Staphylococci and Corynebacterium). Importantly, to validate the behavioral activity of these volatiles, we first assessed landing behavior triggered by human skin volatiles. We demonstrated that landing behavior is gated by the presence of carbon dioxide and L-(+)-lactic acid. This is similar to the combinatorial coding triggering mosquito short range attraction. Repellency behavior to selected skin volatiles and terpenes was tested in the presence of carbon dioxide and L-(+)-lactic acid. In a 2-choice landing behavior context, the skin volatiles 2-and 3-methyl butyric acids reduced mosquito landing by 62.0-81.6% and 87.1-99.6%, respectively. Similarly, the terpene geraniol was capable of reducing mosquito landing behavior by 74.9%. We also tested the potential repellency effects of terpenes in mosquitoes at short-range using a 4-port olfactometer. In these assays, geraniol reduced mosquito attraction (69-78%) to a mixture of key human kairomones carbon dioxide, L-(+)-lactic acid, and ammonia. These findings demonstrate that carbon dioxide and L-(+)-lactic acid change the valence of other skin volatiles towards mosquito landing behavior. Moreover, this study offers candidate odorants to be targeted in a novel strategy to reduce attractants or produce repellents by the human skin microbiota that may curtail mosquito bites, and subsequent mosquito-borne disease. Mosquitoes are one of the biggest threats to human morbidity and mortality around the world due to their exceptional ability to transmit pathogens, including viruses, malaria parasites, and filarial worms. As the number of mosquito vectors resistant to commercial insecticides 1 and vector-borne pathogens gaining resistance to best-in-class drugs 2,3 has increased in recent years 1 , innovative strategies to prevent mosquito bites and pathogen transmission are critical. Ideally, such strategies should protect against the bites of multiple mosquito vectors. Amongst potential new strategies to prevent mosquito bites are the development of safer, affordable, and globally accessible mosquito repellents 4,5. Current strategies aim to disrupt the mosquito chemosensory system using gene editing tools 6-8 and spreading these loss of function mutants into wild populations 9 with some success. With the human skin being the source of numerous attractive odorants, alternative strategies that reduce the production of attractive odorants and/or increase the production of repellents by the human skin might also potentially reduce mosquito bites and pathogen transmission 10,11. Synthetic mosquito repellents such as DEET and picaridin are effective at preventing mosquito bites 12. However, DEET can cause health issues 12 , is unaffordable for widespread use 13 , and requires reapplication within hours 12. In order to find alternative mosquito repellents, chemoinformatics 4 and machine learning approaches 5 have been used to interrogate chemical databases for molecules structurally similar to known repellents. A few

bioRxiv (Cold Spring Harbor Laboratory), Dec 20, 2023

The skin microbiome plays a pivotal role in the production of attractive cues detected by mosquit... more The skin microbiome plays a pivotal role in the production of attractive cues detected by mosquitoes. Here we leveraged recent advances in genetic engineering to significantly reduce the production of L-(+)-lactic acid as a strategy to reduce mosquito attraction to the highly prominent skin commensals Staphylococcus epidermidis and Corynebacterium amycolatum. Engraftment of these engineered bacteria onto the skin of mice reduced mosquito attraction and feeding for up to 11 uninterrupted days, which is considerably longer than the several hours of protection conferred by the leading chemical repellent DEET. Taken together, our findings demonstrate engineering the skin microbiome to reduce attractive volatiles represents an innovative untapped strategy to reduce vector attraction, preventing bites, and pathogen transmission setting the stage for new classes of long-lasting microbiome-based repellent products.

Annual Review of Genetics, Nov 26, 2023

eLife, Jan 21, 2020

Aedes aegypti is the principal mosquito vector for many arboviruses that increasingly infect mill... more Aedes aegypti is the principal mosquito vector for many arboviruses that increasingly infect millions of people every year. With an escalating burden of infections and the relative failure of traditional control methods, the development of innovative control measures has become of paramount importance. The use of gene drives has sparked significant enthusiasm for genetic control of mosquitoes; however, no such system has been developed in Ae. aegypti. To fill this void, here we develop several CRISPR-based split gene drives for use in this vector. With cleavage rates up to 100% and transmission rates as high as 94%, mathematical models predict that these systems could spread anti-pathogen effector genes into wild populations in a safe, confinable and reversible manner appropriate for field trials and effective for controlling disease. These findings could expedite the development of effector-linked gene drives that could safely control wild populations of Ae. aegypti to combat local pathogen transmission.

The resident human skin microbiome is responsible for the production of most of the human scents ... more The resident human skin microbiome is responsible for the production of most of the human scents that are attractive to mosquitoes. Hence, engineering the human skin microbiome to synthesize less of mosquito attractants or produce repellents could potentially reduce bites and prevent the transmission of deadly mosquito-borne pathogens. In order to further characterize the human skin volatilome, we quantified the major volatiles of 39 strains of skin commensals (StaphylococciandCorynebacterium). Importantly, to validate the behavioral activity of these volatiles, we first assessed landing behavior triggered by human skin bacteria volatiles. We demonstrated that this behavioral step is gated by the presence of carbon dioxide and L-(+)-lactic acid, similar to the combinatorial coding triggering short range attraction. Repellency behavior to selected skin volatiles and the geraniol terpene was tested in the presence of carbon dioxide and L-(+)-lactic acid. In a 2-choice landing behavior...

Precision-guided sterile insect technique (pgSIT) is an extremely promising vector control interv... more Precision-guided sterile insect technique (pgSIT) is an extremely promising vector control intervention that can reduce and potentially eliminate the unacceptable malaria burden, particularly in sub-Saharan Africa. Here we explore the cost effectiveness of using this approach in Africa using mathematical modeling and economical analysis. Overall, we find that pgSIT represents a cost-effective and promising approach toA. gambiaecontrol in The Gambia, with the potential to deliver significant economic and social benefits.SummaryPrecision-guided sterile insect technique (pgSIT) is an extremely promising vector control intervention that can reduce and potentially eliminate the unacceptable malaria burden, particularly in sub-Saharan Africa. pgSIT is a safe, innovative, and highly targeted approach to mosquito control that combines the principles of the sterile insect technique (SIT) with advanced state-of-the-art technologies of genetic engineering (Akbari et al. 2023; M. Li et al. 2021...

Insect Molecular Biology, Nov 26, 2019

Title: Methods for the generation of heritable germline mutations in the disease vector Culex qui... more Title: Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using CRISPR/Cas9.

CSH Protocols, Mar 17, 2023

CSH Protocols, Mar 17, 2023

Nature Biotechnology, Dec 15, 2022

Frontiers in Tropical Diseases

Recurring outbreaks of mosquito-borne diseases, like dengue, in the Pacific region represent a ma... more Recurring outbreaks of mosquito-borne diseases, like dengue, in the Pacific region represent a major biosecurity risk to neighboring continents through potential introductions of disease-causing pathogens. Aedes mosquitoes, highly prevalent in this region, are extremely invasive and the predominant vectors of multiple viruses including causing dengue, chikungunya, and Zika. Due to the absence of vaccines for most of these diseases, Aedes control remains a high priority for public health. Currently, international organizations put their efforts into improving mosquito surveillance programs in the Pacific region. Also, a novel biocontrol method using Wolbachia has been tried in the Pacific region to control Aedes mosquito populations. A comprehensive understanding of mosquito biology is needed to assess the risk that mosquitoes might be introduced to neighboring islands in the region and how this might impact arboviral virus transmission. As such, we present a comprehensive review of ...

Frontiers in Tropical Diseases

Genetic-based technologies are emerging as promising tools to support vector population control. ... more Genetic-based technologies are emerging as promising tools to support vector population control. Vectors of human malaria and dengue have been the main focus of these development efforts, but in recent years these technologies have become more flexible and adaptable and may therefore have more wide-ranging applications. Culex quinquefasciatus, for example, is the primary vector of avian malaria in Hawaii and other tropical islands. Avian malaria has led to the extinction of numerous native bird species and many native bird species continue to be threatened as climate change is expanding the range of this mosquito. Genetic-based technologies would be ideal to support avian malaria control as they would offer alternatives to interventions that are difficult to implement in natural areas, such as larval source reduction, and limit the need for chemical insecticides, which can harm beneficial species in these natural areas. This mosquito is also an important vector of human diseases, su...

<p>(A) Southern blot analysis to detect transgene integration sites among transformed <i... more <p>(A) Southern blot analysis to detect transgene integration sites among transformed <i>Ae. aegypti</i>. (B) Southern blot analysis to detect transgene integration sites in Carb109M, Carb109F, Carb109M/GDLS.BC₆, and Carb109F/GDLS.BC₆. Lane 1: HWE; lane 2: Carb109F G₉; lane 3: Carb109M G₉; lane 4: Carb109F/GDLS.BC₆; lane 5: Carb109M/GDLS.BC₆. Total DNA was digested with <i>Pst</i>I. (C) Physical mapping of a transgene integration site in Carb109M mosquitoes. In bold: <i>mariner Mos1</i> TA target sequence motif; in bold and underlined: duplication of the TA target sequence as a consequence of <i>mariner Mos1</i> integration; highlighted in green and blue: partial sequences of the left and right arms of the TE.</p

<p>GDLS, Carb109M, Carb109M/GDLS.BC₅.HZ mosquitoes were challenged w... more <p>GDLS, Carb109M, Carb109M/GDLS.BC₅.HZ mosquitoes were challenged with DENV2-Jamaica1409 (titer in the bloodmeal: >10⁶ pfu/ml). Virus titers of mosquitoes were assessed at 7 and 14 dpi. Each data point represents the virus titer of a single female. Mean values and standard errors are indicated.</p

<p>Initial frequencies (p₀) of the Carb109 transgene were either (A)... more <p>Initial frequencies (p₀) of the Carb109 transgene were either (A) 0.5 (transgenic heterozygote x transgenic heterozygote) for Carb109F/GDLS.BC₁, Carb109M/GDLS.BC₁, Carb109F/GDLS.BC₅, and Carb109M/GDLS.BC₅ or (B) 0.25 (transgenic heterozygote x GDLS) for Carb109F/GDLS.BC₁, Carb109M/GDLS.BC₁, Carb109F/GDLS.BC₅, and Carb109M/GDLS.BC₅. Fifteen lines were established for each of the eight experiments. Proportions of EGFP-expressing offspring were estimated by examining ∼150 larvae from each of the 15 lines over five successive generations (F₁–F₅) of inter-breeding without selection for the transgenic phenotype. Bars around mean proportions represent Bayesian 95% Highest Density Intervals (95% HDI). Proportions showing non-overlapping 95% HDI are credibly different.</p

<p>HWE (control) and Carb109M (G₉, G₁₃, G<... more <p>HWE (control) and Carb109M (G₉, G₁₃, G₁₄) mosquitoes were infected with DENV2 C-932/Acapulco 97, Mex96 Merida, QR94 Quintana Roo, and 14757 Yucatan, representing the Asian 2, Cosmopolitan, American, and Asian-American genotypes, respectively. Mosquitoes received DENV2-containing bloodmeals with the following titers: 1.3×10⁶ pfu/ml (C-932/Acapulco 97), 2.0×10⁵ pfu/ml (Mex96), 4.0×10⁶ pfu/ml (QR94), and 2.0×10⁶ (14757). Virus titers in the mosquitoes were assessed at 7 and 14 dpi. Each data point represents the virus titer of a single female. Mean values and standard errors are indicated.</p

<p>Three replicate crosses (F.1, F.2, F.3) between Carb109F/GDLS.BC5 heterozygotes (200 ind... more <p>Three replicate crosses (F.1, F.2, F.3) between Carb109F/GDLS.BC5 heterozygotes (200 individuals in each cross) and three replicate crosses (M.1, M.2, M.3) between Carb109M/GDLS.BC5 heterozygotes (200 individuals/cross) were performed in separate cages. All resulting wild-type larvae were culled. Remaining transgenic individuals were allowed to inter-mate. This procedure was followed for five generations (F₁–F₅). Results were compared to values expected under Fisher's Selection Model. (A) Observed frequencies of EGFP- expressing Carb109F/GDLS.BC₅ larvae of replicates F.1, F.2, F.3 compared to values predicted under Fisher's Selection model (redline). (B) Observed frequencies of EGFP-expressing Carb109M/GDLS.BC₅ larvae of replicates M.1, M.2, M.3 in comparison to values predicted under Fisher's Selection model (redline). Proportions were compared by estimating the 95% HDI (error bars) with WinBUGS and the Credible Intervals for Proportions script <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002833#pntd.0002833-McCarthy1&quot; target="_blank">[44]</a>.</p

Scientific Reports, Jan 17, 2024

The resident human skin microbiome is responsible for the production of most of the human scents ... more The resident human skin microbiome is responsible for the production of most of the human scents that are attractive to mosquitoes. Hence, engineering the human skin microbiome to synthesize less of mosquito attractants or produce repellents could potentially reduce bites and prevent the transmission of deadly mosquito-borne pathogens. In order to further characterize the human skin volatilome, we quantified the major volatiles of 39 strains of skin commensals (Staphylococci and Corynebacterium). Importantly, to validate the behavioral activity of these volatiles, we first assessed landing behavior triggered by human skin volatiles. We demonstrated that landing behavior is gated by the presence of carbon dioxide and L-(+)-lactic acid. This is similar to the combinatorial coding triggering mosquito short range attraction. Repellency behavior to selected skin volatiles and terpenes was tested in the presence of carbon dioxide and L-(+)-lactic acid. In a 2-choice landing behavior context, the skin volatiles 2-and 3-methyl butyric acids reduced mosquito landing by 62.0-81.6% and 87.1-99.6%, respectively. Similarly, the terpene geraniol was capable of reducing mosquito landing behavior by 74.9%. We also tested the potential repellency effects of terpenes in mosquitoes at short-range using a 4-port olfactometer. In these assays, geraniol reduced mosquito attraction (69-78%) to a mixture of key human kairomones carbon dioxide, L-(+)-lactic acid, and ammonia. These findings demonstrate that carbon dioxide and L-(+)-lactic acid change the valence of other skin volatiles towards mosquito landing behavior. Moreover, this study offers candidate odorants to be targeted in a novel strategy to reduce attractants or produce repellents by the human skin microbiota that may curtail mosquito bites, and subsequent mosquito-borne disease. Mosquitoes are one of the biggest threats to human morbidity and mortality around the world due to their exceptional ability to transmit pathogens, including viruses, malaria parasites, and filarial worms. As the number of mosquito vectors resistant to commercial insecticides 1 and vector-borne pathogens gaining resistance to best-in-class drugs 2,3 has increased in recent years 1 , innovative strategies to prevent mosquito bites and pathogen transmission are critical. Ideally, such strategies should protect against the bites of multiple mosquito vectors. Amongst potential new strategies to prevent mosquito bites are the development of safer, affordable, and globally accessible mosquito repellents 4,5. Current strategies aim to disrupt the mosquito chemosensory system using gene editing tools 6-8 and spreading these loss of function mutants into wild populations 9 with some success. With the human skin being the source of numerous attractive odorants, alternative strategies that reduce the production of attractive odorants and/or increase the production of repellents by the human skin might also potentially reduce mosquito bites and pathogen transmission 10,11. Synthetic mosquito repellents such as DEET and picaridin are effective at preventing mosquito bites 12. However, DEET can cause health issues 12 , is unaffordable for widespread use 13 , and requires reapplication within hours 12. In order to find alternative mosquito repellents, chemoinformatics 4 and machine learning approaches 5 have been used to interrogate chemical databases for molecules structurally similar to known repellents. A few

bioRxiv (Cold Spring Harbor Laboratory), Dec 20, 2023

The skin microbiome plays a pivotal role in the production of attractive cues detected by mosquit... more The skin microbiome plays a pivotal role in the production of attractive cues detected by mosquitoes. Here we leveraged recent advances in genetic engineering to significantly reduce the production of L-(+)-lactic acid as a strategy to reduce mosquito attraction to the highly prominent skin commensals Staphylococcus epidermidis and Corynebacterium amycolatum. Engraftment of these engineered bacteria onto the skin of mice reduced mosquito attraction and feeding for up to 11 uninterrupted days, which is considerably longer than the several hours of protection conferred by the leading chemical repellent DEET. Taken together, our findings demonstrate engineering the skin microbiome to reduce attractive volatiles represents an innovative untapped strategy to reduce vector attraction, preventing bites, and pathogen transmission setting the stage for new classes of long-lasting microbiome-based repellent products.

Annual Review of Genetics, Nov 26, 2023

eLife, Jan 21, 2020

Aedes aegypti is the principal mosquito vector for many arboviruses that increasingly infect mill... more Aedes aegypti is the principal mosquito vector for many arboviruses that increasingly infect millions of people every year. With an escalating burden of infections and the relative failure of traditional control methods, the development of innovative control measures has become of paramount importance. The use of gene drives has sparked significant enthusiasm for genetic control of mosquitoes; however, no such system has been developed in Ae. aegypti. To fill this void, here we develop several CRISPR-based split gene drives for use in this vector. With cleavage rates up to 100% and transmission rates as high as 94%, mathematical models predict that these systems could spread anti-pathogen effector genes into wild populations in a safe, confinable and reversible manner appropriate for field trials and effective for controlling disease. These findings could expedite the development of effector-linked gene drives that could safely control wild populations of Ae. aegypti to combat local pathogen transmission.

The resident human skin microbiome is responsible for the production of most of the human scents ... more The resident human skin microbiome is responsible for the production of most of the human scents that are attractive to mosquitoes. Hence, engineering the human skin microbiome to synthesize less of mosquito attractants or produce repellents could potentially reduce bites and prevent the transmission of deadly mosquito-borne pathogens. In order to further characterize the human skin volatilome, we quantified the major volatiles of 39 strains of skin commensals (StaphylococciandCorynebacterium). Importantly, to validate the behavioral activity of these volatiles, we first assessed landing behavior triggered by human skin bacteria volatiles. We demonstrated that this behavioral step is gated by the presence of carbon dioxide and L-(+)-lactic acid, similar to the combinatorial coding triggering short range attraction. Repellency behavior to selected skin volatiles and the geraniol terpene was tested in the presence of carbon dioxide and L-(+)-lactic acid. In a 2-choice landing behavior...

Precision-guided sterile insect technique (pgSIT) is an extremely promising vector control interv... more Precision-guided sterile insect technique (pgSIT) is an extremely promising vector control intervention that can reduce and potentially eliminate the unacceptable malaria burden, particularly in sub-Saharan Africa. Here we explore the cost effectiveness of using this approach in Africa using mathematical modeling and economical analysis. Overall, we find that pgSIT represents a cost-effective and promising approach toA. gambiaecontrol in The Gambia, with the potential to deliver significant economic and social benefits.SummaryPrecision-guided sterile insect technique (pgSIT) is an extremely promising vector control intervention that can reduce and potentially eliminate the unacceptable malaria burden, particularly in sub-Saharan Africa. pgSIT is a safe, innovative, and highly targeted approach to mosquito control that combines the principles of the sterile insect technique (SIT) with advanced state-of-the-art technologies of genetic engineering (Akbari et al. 2023; M. Li et al. 2021...

Insect Molecular Biology, Nov 26, 2019

Title: Methods for the generation of heritable germline mutations in the disease vector Culex qui... more Title: Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using CRISPR/Cas9.

CSH Protocols, Mar 17, 2023

CSH Protocols, Mar 17, 2023

Nature Biotechnology, Dec 15, 2022

Frontiers in Tropical Diseases

Recurring outbreaks of mosquito-borne diseases, like dengue, in the Pacific region represent a ma... more Recurring outbreaks of mosquito-borne diseases, like dengue, in the Pacific region represent a major biosecurity risk to neighboring continents through potential introductions of disease-causing pathogens. Aedes mosquitoes, highly prevalent in this region, are extremely invasive and the predominant vectors of multiple viruses including causing dengue, chikungunya, and Zika. Due to the absence of vaccines for most of these diseases, Aedes control remains a high priority for public health. Currently, international organizations put their efforts into improving mosquito surveillance programs in the Pacific region. Also, a novel biocontrol method using Wolbachia has been tried in the Pacific region to control Aedes mosquito populations. A comprehensive understanding of mosquito biology is needed to assess the risk that mosquitoes might be introduced to neighboring islands in the region and how this might impact arboviral virus transmission. As such, we present a comprehensive review of ...

Frontiers in Tropical Diseases

Genetic-based technologies are emerging as promising tools to support vector population control. ... more Genetic-based technologies are emerging as promising tools to support vector population control. Vectors of human malaria and dengue have been the main focus of these development efforts, but in recent years these technologies have become more flexible and adaptable and may therefore have more wide-ranging applications. Culex quinquefasciatus, for example, is the primary vector of avian malaria in Hawaii and other tropical islands. Avian malaria has led to the extinction of numerous native bird species and many native bird species continue to be threatened as climate change is expanding the range of this mosquito. Genetic-based technologies would be ideal to support avian malaria control as they would offer alternatives to interventions that are difficult to implement in natural areas, such as larval source reduction, and limit the need for chemical insecticides, which can harm beneficial species in these natural areas. This mosquito is also an important vector of human diseases, su...

<p>(A) Southern blot analysis to detect transgene integration sites among transformed <i... more <p>(A) Southern blot analysis to detect transgene integration sites among transformed <i>Ae. aegypti</i>. (B) Southern blot analysis to detect transgene integration sites in Carb109M, Carb109F, Carb109M/GDLS.BC₆, and Carb109F/GDLS.BC₆. Lane 1: HWE; lane 2: Carb109F G₉; lane 3: Carb109M G₉; lane 4: Carb109F/GDLS.BC₆; lane 5: Carb109M/GDLS.BC₆. Total DNA was digested with <i>Pst</i>I. (C) Physical mapping of a transgene integration site in Carb109M mosquitoes. In bold: <i>mariner Mos1</i> TA target sequence motif; in bold and underlined: duplication of the TA target sequence as a consequence of <i>mariner Mos1</i> integration; highlighted in green and blue: partial sequences of the left and right arms of the TE.</p

<p>GDLS, Carb109M, Carb109M/GDLS.BC₅.HZ mosquitoes were challenged w... more <p>GDLS, Carb109M, Carb109M/GDLS.BC₅.HZ mosquitoes were challenged with DENV2-Jamaica1409 (titer in the bloodmeal: >10⁶ pfu/ml). Virus titers of mosquitoes were assessed at 7 and 14 dpi. Each data point represents the virus titer of a single female. Mean values and standard errors are indicated.</p

<p>Initial frequencies (p₀) of the Carb109 transgene were either (A)... more <p>Initial frequencies (p₀) of the Carb109 transgene were either (A) 0.5 (transgenic heterozygote x transgenic heterozygote) for Carb109F/GDLS.BC₁, Carb109M/GDLS.BC₁, Carb109F/GDLS.BC₅, and Carb109M/GDLS.BC₅ or (B) 0.25 (transgenic heterozygote x GDLS) for Carb109F/GDLS.BC₁, Carb109M/GDLS.BC₁, Carb109F/GDLS.BC₅, and Carb109M/GDLS.BC₅. Fifteen lines were established for each of the eight experiments. Proportions of EGFP-expressing offspring were estimated by examining ∼150 larvae from each of the 15 lines over five successive generations (F₁–F₅) of inter-breeding without selection for the transgenic phenotype. Bars around mean proportions represent Bayesian 95% Highest Density Intervals (95% HDI). Proportions showing non-overlapping 95% HDI are credibly different.</p

<p>HWE (control) and Carb109M (G₉, G₁₃, G<... more <p>HWE (control) and Carb109M (G₉, G₁₃, G₁₄) mosquitoes were infected with DENV2 C-932/Acapulco 97, Mex96 Merida, QR94 Quintana Roo, and 14757 Yucatan, representing the Asian 2, Cosmopolitan, American, and Asian-American genotypes, respectively. Mosquitoes received DENV2-containing bloodmeals with the following titers: 1.3×10⁶ pfu/ml (C-932/Acapulco 97), 2.0×10⁵ pfu/ml (Mex96), 4.0×10⁶ pfu/ml (QR94), and 2.0×10⁶ (14757). Virus titers in the mosquitoes were assessed at 7 and 14 dpi. Each data point represents the virus titer of a single female. Mean values and standard errors are indicated.</p

<p>Three replicate crosses (F.1, F.2, F.3) between Carb109F/GDLS.BC5 heterozygotes (200 ind... more <p>Three replicate crosses (F.1, F.2, F.3) between Carb109F/GDLS.BC5 heterozygotes (200 individuals in each cross) and three replicate crosses (M.1, M.2, M.3) between Carb109M/GDLS.BC5 heterozygotes (200 individuals/cross) were performed in separate cages. All resulting wild-type larvae were culled. Remaining transgenic individuals were allowed to inter-mate. This procedure was followed for five generations (F₁–F₅). Results were compared to values expected under Fisher's Selection Model. (A) Observed frequencies of EGFP- expressing Carb109F/GDLS.BC₅ larvae of replicates F.1, F.2, F.3 compared to values predicted under Fisher's Selection model (redline). (B) Observed frequencies of EGFP-expressing Carb109M/GDLS.BC₅ larvae of replicates M.1, M.2, M.3 in comparison to values predicted under Fisher's Selection model (redline). Proportions were compared by estimating the 95% HDI (error bars) with WinBUGS and the Credible Intervals for Proportions script <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002833#pntd.0002833-McCarthy1&quot; target="_blank">[44]</a>.</p