Delivery Methods for RNAi in Mosquito Larvae

Munawar, Kashif; Alahmed, Azzam; Khalil, Sayed M.S.

doi:10.1093/jisesa/ieaa074

Cited by 20 publications

(13 citation statements)

References 73 publications

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“…RNAi by injection in adult mosquitoes works well for most genes [ 14 , 24 , 25 ]; however, it may not be useful for functional analysis of genes that are highly expressed in larval and pupal stages. Successful RNAi in Ae.…”

Section: Discussionmentioning

confidence: 99%

“…While microinjection in mosquito larvae [ 10 , 11 , 12 ] and pupae [ 13 ] has been shown to work, it is time-consuming and has little field application. An alternative to the microinjection technique is an uptake of dsRNA through ingestion (reviewed in [ 14 ]). In Aedes aegypti, moderate knockdown (30–50%) of the target gene was achieved by simply soaking larvae in a solution of dsRNA for 2 h [ 15 ].…”

Section: Introductionmentioning

confidence: 99%

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RNAi by Soaking Aedes aegypti Pupae in dsRNA

Arshad

Sharma

et al. 2021

Insects

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RNA-interference (RNAi) is a standard technique for functional genomics in adult mosquitoes. However, RNAi in immature, aquatic mosquito stages has been challenging. Several studies have shown successful larval RNAi, usually in combination with a carrier molecule. Except for one study in malaria mosquito, Anopheles gambiae, none of the previous studies has explored RNAi in mosquito pupae. Even in the study that used RNAi in pupae, double stranded RNA (dsRNA) was introduced by microinjection. Here, we describe a successful method by soaking pupae in water containing dsRNA without any carrier or osmotic challenge. The knockdown persisted into adulthood. We expect that this simple procedure will be useful in the functional analysis of genes that highly express in pupae or newly emerged adults.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

RNAi by Soaking Aedes aegypti Pupae in dsRNA

Arshad

Sharma

et al. 2021

Insects

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“…Genes responsible for resistance development in insects (e.g., genes for DDT or pyrethroid resistance) can be identified and used as a target for the development of novel RNAi based insecticides. Several delivery methods including nonmicrobial and microbial are used routinely to induce RNAi in mosquito larvae [199]. Nonmicrobial delivery methods consist of soaking, injection, nanoparticles and dehydration and rehydration.…”

Section: Effective Use Of Plant-based Bioinsecticides In Resistant Mo...mentioning

confidence: 99%

Plant-Based Bioinsecticides for Mosquito Control: Impact on Insecticide Resistance and Disease Transmission

Demirak

Canpolat

2022

Insects

105

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The use of synthetic insecticides has been a solution to reduce mosquito-borne disease transmission for decades. Currently, no single intervention is sufficient to reduce the global disease burden caused by mosquitoes. Problems associated with extensive usage of synthetic compounds have increased substantially which makes mosquito-borne disease elimination and prevention more difficult over the years. Thus, it is crucial that much safer and effective mosquito control strategies are developed. Natural compounds from plants have been efficiently used to fight insect pests for a long time. Plant-based bioinsecticides are now considered a much safer and less toxic alternative to synthetic compounds. Here, we discuss candidate plant-based compounds that show larvicidal, adulticidal, and repellent properties. Our discussion also includes their mode of action and potential impact in mosquito disease transmission and circumvention of resistance. This review improves our knowledge on plant-based bioinsecticides and the potential for the development of state-of-the-art mosquito control strategies.

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“…RNA interference (RNAi) technology refers to the silencing of target gene expression caused by the intervention of double-stranded RNA (dsRNA) [12][13][14][15][16]. This technology has been used in mosquito control recently [17][18][19][20][21]. RNase III-Dicer cleaves dsRNA into small interfering RNA (siRNA) of 20-25 nucleotides in the target cells.…”

Section: Introductionmentioning

confidence: 99%

Effect of marker-free transgenic Chlamydomonas on the control of Aedes mosquito population and on plankton

Fei

Huang

et al. 2023

Parasites Vectors

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Background More than half of the world’s population suffers from epidemic diseases that are spread by mosquitoes. The primary strategy used to stop the spread of mosquito-borne diseases is vector control. Interference RNA (RNAi) is a powerful tool for controlling insect populations and may be less susceptible to insect resistance than other strategies. However, public concerns have been raised because of the transfer of antibiotic resistance marker genes to environmental microorganisms after integration into the recipient genome, thus allowing the pathogen to acquire resistance. Therefore, in the present study, we modified the 3-hydroxykynurenine transaminase (3hkt) and hormone receptor 3 (hr3) RNAi vectors to remove antibiotic resistance marker genes and retain the expression cassette of the inverse repeat sequence of the 3hkt/hr3 target gene. This recombinant microalgal marker-free RNAi insecticide was subsequently added to the suburban water in a simulated-field trial to test its ability to control mosquito population. Methods The expression cassette of the 3hkt/hr3 inverted repeat sequence and a DNA fragment of the argininosuccinate lyase gene without the ampicillin resistance gene were obtained using restriction enzyme digestion and recovery. After the cotransformation of Chlamydomonas, the recombinant algae was then employed to feed Aedes albopictus larvae. Ten and 300 larvae were used in small- and large-scale laboratory Ae.albopictus feeding trials, respectively. Simulated field trials were conducted using Meishe River water that was complemented with recombinant Chlamydomonas. Moreover, the impact of recombinant microalgae on phytoplankton and zooplankton in the released water was explored via high-throughput sequencing. Results The marker-free RNAi-recombinant Chlamydomonas effectively silenced the 3hkt/hr3 target gene, resulting in the inhibition of Ae. albopictus development and also in the high rate of Ae. albopictus larvae mortality in the laboratory and simulated field trials. In addition, the results confirmed that the effect of recombinant Chlamydomonas on plankton in the released water was similar to that of the nontransgenic Chlamydomonas, which could reduce the abundance and species of plankton. Conclusions The marker-free RNAi-recombinant Chlamydomonas are highly lethal to the Ae. albopictus mosquito, and their effect on plankton in released water is similar to that of the nontransgenic algal strains, which reduces the abundance and species of plankton. Thus, marker-free recombinant Chlamydomonas can be used for mosquito biorational control and mosquito-borne disease prevention. Graphical Abstract

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Delivery Methods for RNAi in Mosquito Larvae

Cited by 20 publications

References 73 publications

RNAi by Soaking Aedes aegypti Pupae in dsRNA

RNAi by Soaking Aedes aegypti Pupae in dsRNA

Plant-Based Bioinsecticides for Mosquito Control: Impact on Insecticide Resistance and Disease Transmission

Effect of marker-free transgenic Chlamydomonas on the control of Aedes mosquito population and on plankton

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