Repeat mediated excision of gene drive elements for restoring wild-type populations.

We demonstrate here that single strand annealing (SSA) repair can be co-opted for the precise autocatalytic excision of a drive element. Although SSA is not the predominant form of DNA repair in eukaryotic organisms, we increased the likelihood of its use by engineering direct repeats at sites flanking the drive allele, and then introducing a double-strand DNA break (DSB) at a second endonuclease target site encoded within the drive allele. We have termed this technology Repeat Mediated Excision of a Drive Element (ReMEDE). Incorporation of ReMEDE into the previously described mutagenic chain reaction (MCR) gene drive, targeting the yellow gene of Drosophila melanogaster, replaced drive alleles with wild-type alleles demonstrating proof-of-principle. Although the ReMEDE system requires further research and development, the technology has a number of attractive features as a gene drive mitigation strategy, chief among these the potential to restore a wild-type population without releasing additional transgenic organisms or large-scale environmental engineering efforts.

Engineering a self-eliminating transgene in the yellow fever mosquito, Aedes aegypti.

Promising genetics-based approaches are being developed to reduce or prevent the transmission of mosquito-vectored diseases. Less clear is how such transgenes can be removed from the environment, a concern that is particularly relevant for highly invasive gene drive transgenes. Here, we lay the groundwork for a transgene removal system based on single-strand annealing (SSA), a eukaryotic DNA repair mechanism. An SSA-based rescuer strain (kmoRG ) was engineered to have direct repeat sequences (DRs) in the Aedes aegypti kynurenine 3-monooxygenase (kmo) gene flanking the intervening transgenic cargo genes, DsRED and EGFP. Targeted induction of DNA double-strand breaks (DSBs) in the DsRED transgene successfully triggered complete elimination of the entire cargo from the kmoRG strain, restoring the wild-type kmo gene, and thereby, normal eye pigmentation. Our work establishes the framework for strategies to remove transgene sequences during the evaluation and testing of modified strains for genetics-based mosquito control.

Predictive model for microclimatic temperature and its use in mosquito population modeling.

Mosquitoes transmit several infectious diseases that pose significant threat to human health. Temperature along with other environmental factors at breeding and resting locations play a role in the organismal development and abundance of mosquitoes. Accurate analysis of mosquito population dynamics requires information on microclimatic conditions at breeding and resting locations. In this study, we develop a regression model to characterize microclimatic temperature based on ambient environmental conditions. Data were collected by placing sensor loggers at resting and breeding locations such as storm drains across Houston, TX. Corresponding weather data was obtained from National Oceanic and Atmospheric Administration website. Features extracted from these data sources along with contextual information on location were used to develop a Generalized Linear Model for predicting microclimate temperatures. We also analyzed mosquito population dynamics for Aedes albopictus under ambient and microclimatic conditions using system dynamic (SD) modelling to demonstrate the need for accurate microclimatic temperatures in population models. The microclimate prediction model had an R2 value of ~ 95% and average prediction error of ~ 1.5 °C indicating that microclimate temperatures can be reliably estimated from the ambient environmental conditions. SD model analysis indicates that some microclimates in Texas could result in larger populations of juvenile and adult Aedes albopictus mosquitoes surviving the winter without requiring dormancy.

Making gene drive biodegradable.

Gene drive systems have long been sought to modify mosquito populations and thus combat malaria and dengue. Powerful gene drive systems have been developed in laboratory experiments, but may never be used in practice unless they can be shown to be acceptable through rigorous field-based testing. Such testing is complicated by the anticipated difficulty in removing gene drive transgenes from nature. Here, we consider the inclusion of self-elimination mechanisms into the design of homing-based gene drive transgenes. This approach not only caused the excision of the gene drive transgene, but also generates a transgene-free allele resistant to further action by the gene drive. Strikingly, our models suggest that this mechanism, acting at a modest rate (10%) as part of a single-component system, would be sufficient to cause the rapid reversion of even the most robust homing-based gene drive transgenes, without the need for further remediation. Modelling also suggests that unlike gene drive transgenes themselves, self-eliminating transgene approaches are expected to tolerate substantial rates of failure. Thus, self-elimination technology may permit rigorous field-based testing of gene drives by establishing strict time limits on the existence of gene drive transgenes in nature, rendering them essentially biodegradable. This article is part of the theme issue 'Novel control strategies for mosquito-borne diseases'.

Predicting aquatic development and mortality rates of Aedes aegypti.

Mosquito-borne pathogens continue to be a significant burden within human populations, with Aedes aegypti continuing to spread dengue, chikungunya, and Zika virus throughout the world. Using data from a previously conducted study, a linear regression model was constructed to predict the aquatic development rates based on the average temperature, temperature fluctuation range, and larval density. Additional experiments were conducted with different parameters of average temperature and larval density to validate the model. Using a paired t-test, the model predictions were compared to experimental data and showed that the prediction models were not significantly different for average pupation rate, adult emergence rate, and juvenile mortality rate. The models developed will be useful for modeling and estimating the upper limit of the number of Aedes aegypti in the environment under different temperature, diurnal temperature variations, and larval densities.

Framework for Infectious Disease Analysis: A comprehensive and integrative multi-modeling approach to disease prediction and management.

The impact of infectious disease on human populations is a function of many factors including environmental conditions, vector dynamics, transmission mechanics, social and cultural behaviors, and public policy. A comprehensive framework for disease management must fully connect the complete disease lifecycle, including emergence from reservoir populations, zoonotic vector transmission, and impact on human societies. The Framework for Infectious Disease Analysis is a software environment and conceptual architecture for data integration, situational awareness, visualization, prediction, and intervention assessment. Framework for Infectious Disease Analysis automatically collects biosurveillance data using natural language processing, integrates structured and unstructured data from multiple sources, applies advanced machine learning, and uses multi-modeling for analyzing disease dynamics and testing interventions in complex, heterogeneous populations. In the illustrative case studies, natural language processing from social media, news feeds, and websites was used for information extraction, biosurveillance, and situation awareness. Classification machine learning algorithms (support vector machines, random forests, and boosting) were used for disease predictions.

Impacts of diurnal temperature and larval density on aquatic development of Aedes aegypti.

The increasing range of Aedes aegypti, vector for Zika, dengue, chikungunya, and other viruses, has brought attention to the need to understand the population and transmission dynamics of this mosquito. It is well understood that environmental factors and breeding site characteristics play a role in organismal development and the potential to transmit pathogens. In this study, we observe the impact of larval density in combination with diurnal temperature on the time to pupation, emergence, and mortality of Aedes aegypti. Experiments were conducted at two diurnal temperature ranges based on 10 years of historical temperatures of Houston, Texas (21-32°C and 26.5-37.5°C). Experiments at constant temperatures (26.5°C, 32°C) were also conducted for comparison. At each temperature setting, five larval densities were observed (0.2, 1, 2, 4, 5 larvae per mL of water). Data collected shows significant differences in time to first pupation, time of first emergence, maximum rate of pupation, time of maximum rate of pupation, maximum rate of emergence, time of maximum rate of emergence, final average proportion of adult emergence, and average proportion of larval mortality. Further, data indicates a significant interactive effect between temperature fluctuation and larval density on these measures. Thus, wild population estimates should account for temperature fluctuations, larval density, and their interaction in low-volume containers.

Influence of Processing Techniques on Microstructure and Mechanical Properties of a Biodegradable Mg-3Zn-2Ca Alloy.

New Mg-3Zn-2Ca magnesium alloy was prepared using different processing techniques: gravity casting as well as squeeze casting in liquid and semisolid states. Materials were further thermally treated; thermal treatment of the gravity cast alloy was additionally combined with the equal channel angular pressing (ECAP). Alloy processed by the squeeze casting in liquid as well as in semisolid state exhibit improved plasticity; the ECAP processing positively influenced both the tensile and compressive characteristics of the alloy. Applied heat treatment influenced the distribution and chemical composition of present intermetallic phases. Influence of particular processing techniques, heat treatment, and intermetallic phase distribution is thoroughly discussed in relation to mechanical behavior of presented alloys.