Books and Chapters of Books
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Item Engineering RNA interference-based dengue virus resistance in the mosquito vector aedes aegypti: The current status and future directions(Springer, 2021) Denipitiyage, S.D.; Gunawardene, Y.I.N.S.; Federico, Z.; Dassanayake, R.S.Dengue is an acute, febrile disease caused by the dengue viruses (DENV) comprising four serotypes and transmitted by the mosquito vector Ae. aegypti. DENV are single-stranded, positive-sense RNA viruses of the family Flaviviridae. Dengue is declared as a current significant challenge in the Southeast Asia, imposing growing burden on infected populations. To date, dengue control has mostly relied on vector control strategies which have largely become ineffective. There is, therefore, an urgent need for novel vector control strategies. Development of genetically modified mosquito vectors to manipulate disease-vectoring populations has gathered increased interest in recent time. RNAi-mediated viral resistance contributes to the suppression of viruses, including DENV in the mosquito vector Ae. aegypti. With recent advances in the field of molecular biology, we and other scientists are continuing to engineer genes that confer virus resistance to reduce transmission rates of DENV and introducing these genes into the mosquito genome. Even though scientists successfully generated mosquito refractory to DENV2–4, no mosquito refractory to all four serotypes has been developed to date. This limitation can be overcome by systematic analysis of the molecular mechanisms of RNAi in the mosquito vector Ae. aegypti. An enhanced understanding of RNAi function in the mosquito vector Ae. aegypti will facilitate the application of RNAi to control the transmission of the dengue disease in the future. Here, based on current understanding of the RNAi, we discuss the mechanisms of RNAi in the mosquito vector Ae. aegypti. We also provide guidelines for optimal design of RNAi experiments in Ae. aegypti with the possible risks associated with them along with proposed solutions.Item Advances in Aedes mosquito vector control strategies using CRISPR/Cas9(Springer, 2021) Wickramasinghe, P.D.S.U.; Silva, G.N.; Gunawardene, Y.I.N.S.; Dassanayake, R.S.Advancements in genetic engineering have resulted in the development of mosquitoes with impaired vector competence, thereby limiting acquisition and transmission of pathogens. The main dengue (DENV) vector, Aedes aegypti, is an invasive species that have spread unwittingly across the world as a result of human trade and travel. The Ae. aegypti mosquito species has spread across tropical and subtropical regions, with higher presence in urban regions where rapid breeding patterns have shown in artificial containers. Identification of and treating an adequate number of mosquito breeding sites as a control measure have been done for the past couple of years, and yet improvement is far from the expectations, even with well-funded and well-organized initiatives. In order to stop the pathogen transmission, genetically modified mosquitoes (GMM) needs to be created and released. Despite many Aedes-related achievements, GMM creation has been challenging. The spread of particular genetic elements that impair vector competence, trigger deleterious recessive mutations, or skew a population's sex ratio can be used to prevent the spread of vector disease, or eradicate invasive organisms in a species-specific and eco-friendly manner. In recent years, genome editing strategies have evolved to make use of a variety of nucleases, ranging from sequence-specific zinc finger nucleases to modular TALENs (transcription activator-like effector nucleases) and most recently, RNA-guided nucleases adapted from bacterial adaptive immune systems, dubbed CRISPR/Cas (clustered regularly interspaced palindromic repeats/CRISPR associated systems). By combining these methods, a new era in gene editing had emerged. Generally, both of these gene editing technologies utilize sequence-specific nucleases to generate double-stranded DNA breaks (or nicks) in the target sequence, resulting in desired DNA modifications using endogenous DNA repair mechanisms. Since cells with DNA lesions are unable to divide further, the nuclease-generated strand breaks must be rapidly repaired by the cell to maintain the viability. CRISPR/Cas has been widely accepted for use in a variety of organisms, including insect species, with only minor optimization steps needed thus far. CRISPR/Cas9 technology transformed the process of engineering nucleases capable of cleaving complex genomic sequences. A complementary guide RNA (gRNA) directs the Cas9 endonuclease's operation to the specific DNA target site, enabling the editing of virtually any DNA sequence without complex protein engineering and selection procedures. Apart from genome editing, the specificity and flexibility of the CRISPR/Cas9 method enables unprecedented rapid development of genetically modified organisms with mutation systems for disease vector insect control. The stability and expression of the gene construct generated by CRISPR/Cas9 or any other method must be addressed before GMM are released, in order to make sure that pathogen transmission and formulation are interrupted robustly and completely. Spreading foreign antipathogen genes through gene drive strategies among wild mosquito populations strengthens the case for a more streamlined approach. Major fields that must be adequately assessed include risk evaluation and management, conducting studies to ensure human and environmental protection, developing effective control strategies built on comprehensive gene-driving systems, and adequately addressing the ethical, legal, and social consequences of GMM release. Although GMM is theoretically feasible as a disease control method, field releases should be made only when strong scientific evidence of human and environmental protection and effectiveness are presented, and public acceptance is addressed appropriately. This chapter discusses the diverse technological advances in generating Ae. aegypti mosquitoes which are resistant to dengue virus (DENV) and other diseases, as well as the biosafety and risk assessment of these procedures. Additionally, the chapter outlines a convincing path forward for developing successful genetic-based DENV control strategies based on CRISPR/Cas9, which could be expanded to control other arboviruses while maintaining biosafety.Item Genetic improvements to the sterile insect technique (SIT) for the control of mosquito population(Springer, 2021) Dilani, P.V.D.; Gunawardene, Y.I.N.S.; Dassanayake, R.S.Mosquito-borne diseases are becoming a major health problem worldwide. At present, the principal method of controlling these diseases entirely depends on the mosquito vector control strategies. However, traditional control methods which are focussed on reducing mosquito populations through environmental management and the application of insecticides are largely ineffective. Hence, various control methods, including the release of sterile insect technique (SIT), have been proposed for the reduction of the mosquito population. As a species-specific control strategy, SIT offers considerable environmental benefits and a chemical-free option for insect control. However, the application of the SIT to mosquito control consistently suffered from lack of efficient sexing system, high fitness cost and operational difficulty in ionizing radiation, density-dependent nature of the target mosquito population and various other technical issues. The intervention of genetic engineering has led to several improvements in the operation or security of SIT programmes. The advent of mosquito transgenesis has paved the way for novel approaches in mosquito control. One possibility is a release of insects carrying dominant lethal (RIDL) strategy by engineering self-limiting gene, which offers solutions for many drawbacks of traditional SIT by providing genetic sterilization, genetic sexing, genetic containment and provision of genetic markers while maintaining its environmentally benign and species-specific utility. The success of this strategy often depends on how genetic modification affects the fitness of the mosquitoes. With several improvements and modifications allowing minimum fitness load, RIDL is now available for a wide range of mosquitoes such as Aedes aegypti, Aedes albopictus and Anopheles stephensi with field-testing possibilities. However, with solid epidemiological evidence and community support, widespread implementation of these strategies might reverse the current alarming global mosquito vector-borne diseases.Item Genome, gene expression and regulation in Prokaryotes(Author, 2009) Gunawardene, Y.I.N.S.No abstract availableItem Methyl Farnesoate in Crustacean Endocrine control(Author, 2009) Gunawardene, Y.I.N.S.No abstract availableItem Item Bioinformatics and DNA micro-arrays in post genomic analysis(Author, 2009) Dassanayake, R.S.; Gunawardene, Y.I.N.S.No abstract available