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CRISPR-Cas9-mediated genetic modifications in mosquitoes. The figure shows two kinds of mutants such as knockout and knock-in mutants produced by CRISPR/Cas9-mediated genome editing. It could be crossed with a wild type mosquito in a gene drive system to disrupt the particular activity/functions of the vector mosquito and population suppression that leads to disease eradication. The altered genes could be preferentially inherited by all offsprings when crossed with a wild type mosquito. A Gene drive technology would quickly spread the altered gene in the target mosquito population with nearly 100% chance. Vg -vestigial gene.
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Mosquitoes pose a major threat to human health as they spread many deadly diseases like malaria, dengue, chikungunya, filariasis, Japanese encephalitis and Zika. Identification and use of novel molecular tools are essential to combat the spread of vector borne diseases. Genome editing tools have been used for the precise alterations of the gene of...
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Genetic modification has long provided an approach for reverse genetics i.e. analyzing gene function and linking DNA sequence to phenotype. However, traditional genome modification technologies have not kept pace with the soaring progress of the genome sequencing era, due to their inefficiency, time-consuming and labor-intensive features. Recently,...
Gene editing tools like TALENs, ZFNs and Crispr/Cas now offer unprecedented opportunities for targeted genetic manipulations in virtually all species. Most of the recent research in this area has concentrated on manipulation of the genome in isolated cells, which then give rise to transgenic animals or modified stem cell lines. Much less is known a...
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Citations
... Mosquitoes are well-known carriers of deadly diseases that pose a significant threat to human health. Conditions such as filariasis, chikungunya, dengue, zika, and malaria have caused immense suffering and loss of life (Gabrieli et al., 2014;Reegan et al., 2017). Traditionally, synthetic insecticides have been employed to combat disease-carrying mosquitoes. ...
The field of life science research has undergone a revolution thanks to the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) and CRISPR-associated protein (CRISPR/Cas) system, which provides a multitude of opportunities for modifying, identifying, visualising, and annotating particular DNA or RNA sequences in diverse organisms. In this technique, foreign DNA pieces, known as spacers, are inserted into CRISPR cassettes. These spacers are then transcribed into CRISPR arrays and processed to produce guide RNA (gRNA). The Cas proteins that the CRISPR arrays encode serve as the enzymatic machinery required to obtain new spacers that specifically target invasive genetic elements. Several Cas proteins, such as Cas9, Cas12, Cas13, and Cas14, have been used to create novel tools for genome engineering due to their programmable sequence specificity. The ability to manipulate and edit nucleic acid sequences in living cells from a wide variety of organisms has been made possible by these Cas variants, which have greatly advanced genetic research and the CRISPR/Cas tool. The CRISPR Cas-9 technology has applications in many areas of entomology, including the genetics of honeybees and plants that produce insecticidal compounds. CRISPR/Cas9 technology has transformed entomology by providing precise tools for gene editing and genetic manipulation in insects. This has enabled advancements in fundamental research, disease vector control, and pest management, with the potential to reduce the environmental and economic impact of insect pests in agriculture and public health.
... TALEN and ZFNs use protein-DNA interactions to identify DNA sequences, but these methods have definite drawbacks, made plasmid construction a challenge, and are also very expensive (Kumar et al. 2015;Soda et al. 2018). TALEN is less costly, easier to construct than ZFN technology, and more specific to the target sequence (Reegan et al. 2016). However, compared to ZFN and TALEN, the CRISPR system is currently considered the most effective, affordable, and flexible cost-effective genome editing technique Chang et al. 2016). ...
Main conclusion
This review provides a comprehensive overview of the CRISPR/Cas9 technique and the research areas of this gene editing tool in improving wheat quality.
Abstract
Wheat (Triticum aestivum L.), the basic nutrition for most of the human population, contributes 20% of the daily energy needed because of its, carbohydrate, essential amino acids, minerals, protein, and vitamin content. Wheat varieties that produce high yields and have enhanced nutritional quality will be required to fulfill future demands. Hexaploid wheat has A, B, and D genomes and includes three like but not identical copies of genes that influence important yield and quality. CRISPR/Cas9, which allows multiplex genome editing provides major opportunities in genome editing studies of plants, especially complicated genomes such as wheat. In this overview, we discuss the CRISPR/Cas9 technique, which is credited with bringing about a paradigm shift in genome editing studies. We also provide a summary of recent research utilizing CRISPR/Cas9 to investigate yield, quality, resistance to biotic/abiotic stress, and hybrid seed production. In addition, we provide a synopsis of the laboratory experience-based solution alternatives as well as the potential obstacles for wheat CRISPR studies. Although wheat’s extensive genome and complicated polyploid structure previously slowed wheat genetic engineering and breeding progress, effective CRISPR/Cas9 systems are now successfully used to boost wheat development.
... The genome editing tool like clustered, regularly inter-spaced, short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) creates an novel option to study insect genetics. Several studies have confirmed the utilizing of the CRISPR/Cas9 system to control vector mosquitoes (reviewed in Hillary et al., 2020;Hillary and Ceasar, 2021;Reegan et al., 2016). Employing these genome-editing approaches in combating disease spreading vector mosquitoes will open new avenues in the upcoming future. ...
Alternative ecofriendly products to control vector borne diseases are urgently needed to combat mosquito re-lated diseases. New modes of action, cost effectiveness, bio-degradability, and target specificity are some de-sirable attributes for these alternative products. Studies on macrofungi have become increasingly important forresearchers in the current scenario. Macrofungi are widespread and are found all over the world. Basidiomycetes(mushrooms) are found mostly in warm and humid climatic conditions. No specific review is available on thebasidiomycetes for mosquito control till 2022. A number of online libraries including PubMed, Scifinder, GoogleScholar and Web of Science were used in searching for information on mushrooms for mosquito control. Naturalantibiotics are abundant in basidiomycetes; they have shown antimicrobial, anticancer, antioxidant and other biological activities. This review summarises the potential role of mushrooms as promising and effective mos-quito control agents
(PDF) Mushrooms (Basidiomycetes) as Source of Mosquito Control Agents. Available from: https://www.researchgate.net/publication/371381440_Mushrooms_Basidiomycetes_as_Source_of_Mosquito_Control_Agents [accessed Jun 14 2023].
... 1 It is a dominant outdoor breeding species capable of spreading dengue, chikungunya, and Zika virus. [2][3][4][5] Being an important diseasecausing agent, it also creates irritation and biting nuisance affecting the daily life of humans. 6 Factors like climate change and unplanned urbanisation have led to frequent exposure of humans to this mosquito. ...
Introduction: The emergence of resistance development in mosquitoes is a huge challenge for successful vector control programmes. Insecticides used in vector control mainly target the eradication of the vector population but often neglect their residual and long-time effect on the surviving population. Methods: In this study, Aedes albopictus were selected with sublethal doses of lambda-cyhalothrin and its effects on their life parameters and major detoxifying enzymes were analysed. Results: In the twelfth generation of the resistant population, there was a 101-fold increase in resistance ratio values of LC50. In addition, larval development time prolonged for 2 days and hatchability and fecundity were reduced by 52% and 93%, respectively when compared to the control group. Moreover, longevity was shortened by 9.43 days in females and 2.18 days in males. Significant changes in the activity of major detoxifying enzymes were observed where monooxygenase level was highest which indicates its role in lambda-cyhalothrin degradation leading to resistance, followed by both β-esterase and GST. Conclusion: The findings of the current study can be helpful in integrated mosquito management (IMM), where based on their life history traits, the vulnerable stage of vectors can be targeted for effective resistance management.
... Unlike men, the female gene contains one exon (exon 5), the sequence of which is very conserved in all mosquitoes. By blocking the 4-exon 5 junction, the CRISPR-Cas9 system prevents AgdsxF production without affecting male development and reproduction.Females with destroyed allele homozygous showed bisexual phenotype and complete infertility [11]. ...
Malaria, which is primarily transmitted by female mosquitoes, is increasing recognized as a global public health problem. Since the number of malaria cases remains high, with more than 200 million cases globally in 2019 alone, the development of new drugs and corresponding prevention and control measures are in urgent need. With the development of CRISPR/Cas9 technology in recent years, gene editing technology plays an increasingly significant role in mosquito control. A new generation of gene drive based on CRISPR/Cas9 has been applied into improving the heritability of specific genes in mosquito populations, which can be exploited to control mosquito and combat diseases transmitted by mosquito. However, this technology still faces challenges of ecological environment safety, ethical safety and technical effectiveness. This review introduces the current research status of gene drive systems for mosquito control using the CRISPR/Cas 9 system, and summarizes the current problems and solutions in this research field.
... As a result, the molecular and neuronal mechanisms of host choice have received increased attention in the last few years, driven by technological advancements in, e.g. next generation sequencing and gene editing tools for mosquitoes (e.g. reviewed in Chen et al., 2021;Reegan et al., 2016;Riabinina et al., 2016;Zhao et al., 2021). ...
... In the past few years, synthetic pesticides have been used to control mosquitoes. However, synthetic pesticides can cause contamination, kill many beneficial insects, and lead to the development of resistant-types after long-term use [3]. positive results, and misdiagnosis. ...
... A major advance in this area is the discovery of sgRNA. It was originally used in combination with Cas9 and made in vitro cuts at various DNA sites [3]. ...
Tropical diseases (TDs) are among the leading cause of mortality and fatality globally. The emergence and reemergence of TDs continue to challenge healthcare system. Several tropical diseases such as yellow fever, tuberculosis, cholera, Ebola, HIV, rotavirus, dengue, and malaria outbreaks have led to endemics and epidemics around the world, resulting in millions of deaths. The increase in climate change, migration and urbanization, overcrowding, and other factors continue to increase the spread of TDs. More cases of TDs are recorded as a result of substandard health care systems and lack of access to clean water and food. Early diagnosis of these diseases is crucial for treatment and control. Despite the advancement and development of numerous diagnosis assays, the healthcare system is still hindered by many challenges which include low sensitivity, specificity, the need of trained pathologists, the use of chemicals and a lack of point of care (POC) diagnostic. In order to address these issues, scientists have adopted the use of CRISPR/Cas systems which are gene editing technologies that mimic bacterial immune pathways. Recent advances in CRISPR-based biotechnology have significantly expanded the development of biomolecular sensors for diagnosing diseases and understanding cellular signaling pathways. The CRISPR/Cas strategy plays an excellent role in the field of biosensors. The latest developments are evolving with the specific use of CRISPR, which aims for a fast and accurate sensor system. Thus, the aim of this review is to provide concise knowledge on TDs associated with mosquitoes in terms of pathology and epidemiology as well as background knowledge on CRISPR in prokaryotes and eukaryotes. Moreover, the study overviews the application of the CRISPR/Cas system for detection of TDs associated with mosquitoes.
... Advances in synthetic biology provides a new potential tool for population control of invasive species, which is known as genetic biocontrol. Research is currently underway to develop genetic biocontrol systems for multiple species including disease carrying mosquitoes [3,39,62] and mice. [38] In the cane toad the fundamental synthetic biology tools necessary to make genetic biocontrol a possible option in the future are being developed with the success generation gene edited cane toads. ...
Synthetic biology [synbio] applications have the potential to assist in addressing significant global health and environmental challenges. Australian research institutes are investing in formative research to develop synbio technologies capable of meeting these challenges. Alongside the laboratory research, investigating the broader social, institutional and ethical considerations that synbio presents has been a priority. We conducted targeted qualitative research to uncover the barriers and opportunities for a range of multisectoral stakeholders identified as potential end‐users of the science under development. The research provides insights into the research implementation environment for three synthetic biology applications: (1) gene editing cane toads (Rhinella marina) to reduce their environmental impact; (2) engineering bacteriophages to combat antimicrobial resistance in humans and; (3) engineering microbes to improve biomining efficiency in the mining industry. In‐depth interviews (N = 23) with government, research and civil society representatives revealed key challenges in the impact pathway for each application. The strongest themes uncovered during interviews related to perceived negative public attitudes towards genetic technologies, a lack of investment in critical research infrastructure, unclear regulatory pathways and the presence of a strong social and environmental imperative underpinning technology development. These findings reveal specific entry points for further engagement with the most immediate end‐users of synbio. Separate from research on public attitudes to synbio, the cases highlight the various hurdles to achieving research impact, according to experts who will likely use, approve or invest in these applications in the future. The themes uncovered inform avenues for strengthening engagement and research coordination in Australia and elsewhere. This article is protected by copyright. All rights reserved
... Fuelled by recent advancement in the genetic toolkit of mosquitoes (e.g., Reegan et al., 2016;Matinyan and Potter, 2022), comparative transcriptomic studies identified several chemosensory genes, encoding for ORs, IRs and OBPs, to be differentially expressed in closely related species with divergent host preference (Rinker et al., 2013;Neafsey et al., 2015;Athrey et al., 2017). Furthermore, by knocking out the OR co-receptor gene, orco, DeGennaro et al. (2013) demonstrated that the OR-pathway is necessary for host discrimination. ...
... Despite reporting negative results, this study (paper III) is among the first to investigate the mechanism by which a single tuning OR can affect mosquito host-seeking behaviour. With the recent progress in the development of genetic tools in mosquitoes, the field is now ready to advance one step further and establish causality, not just correlation, between mosquito behaviour and, e.g., regulation of chemosensory genes (Reegan et al., 2016;Zhao and McBride, 2020;Matinyan and Potter, 2022). ...
The majority of the world’s population is at risk of one or more mosquito-borne diseases that are transmitted by blood-feeding female mosquitoes, affecting both human health and economic development. Especially Anopheles gambiae, the principal malaria vector, and Aedes aegypti, the vector of dengue and yellow fever, are of primary concern due to their strong specialisation on human hosts, and the high number of casualties caused by the pathogens they transmit. Host seeking and discrimination are crucial for disease transmission, and are predominantly mediated by olfaction. Using a wind tunnel system and a custom analysis pipeline, this thesis confirms that the two mosquito species use volatile host cues, derived from breath and body, differentially, as carbon dioxide on its own drives host seeking in Ae. aegypti, but not in An. gambiae (paper I). To discriminate between host and nonhost species (paper V), Ae. aegypti encode human identity by the relative activation of two glomeruli within the antennal lobe, the primary olfactory centre, of which one is tuned to long-chain aldehydes enriched in human odour. A synthetic blend mimicking the glomerular activation elicited host seeking in Ae. aegypti (paper II). Next to preferring human over non-human hosts, Ae. aegypti also prefer some human individuals to others, which was demonstrated to be affected by the ABO blood type and pregnancy or menstrual cycle phase. Analysis of the volatiles associated with individual volunteers, identified 1-octen-3-ol to be significantly associated with very high attractiveness (paper III). The molecular regulation of host seeking acquisition during An. gambiae female adult maturation was independent of odorant receptor gene AgamOR39 expression (paper IV). The results presented in this thesis contribute to the understanding of mosquito host seeking and discrimination from multiple perspectives, which is a prerequisite to ultimately develop novel tools for mosquito monitoring and control.
... Gene-editing technologies have expanded possibilities of vector control by allowing researchers to edit target genes, analyze gene functions, and even gene expression. 6 There are currently at least 12 classes of genetic technologies that may be used on disease vectors, including zinc finger nucleases, 7 transcription activator-like effector nucleases, 8 and CRISPR-Cas9. 9 CRISPR-Cas9 technology is a genome-editing tool rising in popularity due to its versatility, low cost, specificity, and simplicity. ...
... An increase in drive-resistant alleles in natural populations, especially after mating of transgenic mosquitoes and wild-type mosquitoes, could lead to undesirable downstream effects. 6 ...
