Synthetic miRNAs induce dual arboviral-resistance phenotypes in the vector mosquito Aedes aegypti

Yen, Pei‐Shi; James, Anthony A.; Li, Jian-Chiuan; Chen, Chun Hong; Failloux, Anna‐Bella

doi:10.1038/s42003-017-0011-5

Cited by 59 publications

(67 citation statements)

References 69 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The antiviral effectors developed in Ae. aegypti to date rely upon small RNA pathway-directed targeting of viral genomes (26,(61)(62)(63)(64); however, these antiviral transgenes are not easily multiplexed and the diversity of arboviruses transmitted by this vector as well as the high mutation rate of these viruses (65) makes their practicality in the field tenuous. Additionally, long-term studies of viral resistance to these antiviral effectors are limited, so their long-term effectiveness is unknown.…”

Section: Discussionmentioning

confidence: 99%

“…This process converts wild-type alleles into drive alleles in heterozygotes, thereby forcing super-mendelian inheritance of the drive into subsequent generations, irrespective of the fitness cost to the organism. Theoretically, this inheritance scheme can spread the drive and any linked anti-pathogen "cargo genes," to fixation in a population in a short time frame (26)(27)(28), even with modest introduction frequencies (19). Importantly, if these drives are linked with effective anti-pathogen cargo genes, they could deliver efficient and cost-effective vector control (19).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Development of a Confinable Gene-Drive System in the Human Disease Vector,Aedes aegypti

Li¹,

Yang²,

Kandul³

et al. 2019

Preprint

View full text Add to dashboard Cite

Aedes aegypti, the principal mosquito vector for many arboviruses that causes yellow fever, dengue, Zika, and chikungunya, increasingly infects 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 recently sparked significant enthusiasm for the genetic control of mosquito populations, however no such system has been developed in Ae. aegypti. To fill this void and demonstrate efficacy in Ae. aegypti, 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%, 2 mathematical models predict that these systems could spread anti-pathogen effector genes into wild Ae. aegypti populations in a safe, confinable and reversible manner appropriate for field trials and effective for controlling disease. These findings could expedite the development of effectorlinked gene drives that could safely control wild populations of Ae. aegypti to combat local pathogen transmission. Significance StatementAe. aegypti is a globally distributed arbovirus vector spreading deadly pathogens to millions of people annually. Current control methods are inadequate and therefore new technologies need to be innovated and implemented. With the aim of providing new tools for controlling this pest, here we engineered and tested several split gene drives in this species. These drives functioned at very high efficiency and may provide a tool to fill the void in controlling this vector. Taken together, our results provide compelling path forward for the feasibility of future effector-linked split-drive technologies that can contribute to the safe, sustained control and potentially the elimination of pathogens transmitted by this species.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Development of a Confinable Gene-Drive System in the Human Disease Vector,Aedes aegypti

Li¹,

Yang²,

Kandul³

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…Numerous RNA viruses of global medical importance, such as dengue, Zika, and chikungunya virus, are transmitted by one species of Aedes mosquito. Engineering this mosquito population with virus resistance may be a tool to reduce disease transmission [11]; however, no current technologies have successfully targeted all of these viruses simultaneously [12][13][14][15][16]. RNAtargeting CRISPR systems may provide a platform to reduce the spread of mosquito-borne viruses by targeting viral RNA genomes in a programmable manner.…”

Section: Introductionmentioning

confidence: 99%

Programmable RNA Targeting using CasRx in Flies

Buchman¹,

Brogan²,

Sun³

et al. 2020

Preprint

View full text Add to dashboard Cite

CRISPR-Cas genome editing technologies have revolutionized the fields of functional genetics and genome engineering, but with the recent discovery and optimization of RNA-targeting Cas ribonucleases, we may soon see a similar revolution in the study of RNA function and transcriptome engineering. However, to date, successful proof-of-principle for Cas ribonuclease RNA targeting in eukaryotic systems has been limited. Only recently has successful modification of RNA expression by a Cas ribonuclease been demonstrated in animal embryos. This previous work, however, did not evaluate endogenous expression of Cas ribonucleases and only focused on Cas ribonuclease function in early developmental stages. A more comprehensive evaluation of this technology is needed to assess its potential impact in the field. Here we report on our efforts to develop a programmable platform for RNA-targeting using a Cas ribonuclease, CasRx, in the model organism Drosophila melanogaster. By genetically encoding CasRx in flies, we demonstrate moderate transcript targeting of known phenotypic genes in addition to unexpected toxicity and lethality. We also report on the off-target effects following on-target transcript cleavage by CasRx. Taken together, our results present the current state and limitations of a genetically encoded programmable RNA-targeting Cas system in Drosophila melanogaster, paving the way for future optimization of the system.

show abstract

“…To generate a "cargo" gene that can confer resistance to ZIKV, we implemented a synthetic miRNA-based approach, since such an approach has been previously demonstrated to generate virus-resistance phenotypes in a number of contexts (e.g., [28][29][30] ), including mosquitoes 31 . We engineered a piggyBac vector comprising a polycistronic cluster of eight ZIKV-targeting synthetic miRNAs (anti-ZIKV transgene) under control of the Ae.…”

Section: Engineering Zikv-resistant Mosquitoesmentioning

confidence: 99%

Engineered resistance to Zika virus in transgenicAe. aegyptiexpressing a polycistronic cluster of synthetic miRNAs

Buchman

Gamez

et al. 2018

Preprint

Self Cite

View full text Add to dashboard Cite

One Sentence Summary:Here we describe the generation of Ae. aegypti mosquitoes that are engineered to confer reduced vector competence to Zika virus (ZIKV) and we discuss how such engineering approach can be used to combat the major health burden of ZIKV and potentially other arboviruses in the future. Abstract:Recent Zika virus (ZIKV) outbreaks have highlighted the necessity for development of novel vector control strategies to combat arboviral transmission, including genetic versions of the sterile insect technique, artificial infection with Wolbachia to reduce population size and/or vectoring competency, and gene drive based methods. Here, we describe the development of mosquitoes synthetically engineered to impede vector competence to ZIKV. We demonstrate that a polycistronic cluster of engineered microRNAs (miRNAs) targeting ZIKV is expressed and fully processed following a blood meal in Ae. aegypti , ensuring the formation of mature synthetic miRNAs in the midgut where ZIKV resides in the early stages of infection. Critically, we demonstrate that engineered Ae. aegypti mosquitoes harboring the anti-ZIKV transgene have significantly reduced viral infection, dissemination, and transmission rates of ZIKV. Taken together, these compelling results provide a promising path forward for development of effective genetic-based ZIKV control strategies, which could potentially be extended to curtail other arboviruses. MainSince being introduced into the Americas, ZIKV -a mosquito-borne flavivirus -has spread rapidly, causing hundreds of thousands of cases of ZIKV infection 1 . Although most cases remain asymptomatic, infection during pregnancy has been associated with severe congenital abnormalities and pregnancy loss, presenting an unprecedented health threat with long-term consequences 2 . This prompted the World Health Organization to declare ZIKV a Public Health Emergency of International Concern in 2016 1,2 . Currently, there are no clinically approved vaccines to prevent ZIKV and no effective treatment options for infected individuals; thus, vector control remains essential in curtailing the ZIKV epidemic. Like dengue virus (DENV) and chikungunya virus (CHIKV), ZIKV is transmitted primarily by Aedes mosquitoes, which are expanding their habitable range due to urbanization, climate change, and global trade 3 . Current methods of vector control, including removal of standing water and use of insecticides, have not been entirely effective in the fight against the spread of Aedes mosquitoes 3 . Therefore, new game-changing innovative vector control strategies, including those utilizing genetically engineered mosquitoes 4 , are urgently needed to combat the spread of ZIKV and other Aedes -vectored diseases worldwide.Employment of genetically modified (or otherwise altered) insects to manipulate disease-vectoring populations was first proposed decades ago 5 , and due in part to enabling technological advances, has garnered increased interest in recent years 4 . In fact, several strategies for genetic-based vector contr...

show abstract

Synthetic miRNAs induce dual arboviral-resistance phenotypes in the vector mosquito Aedes aegypti

Cited by 59 publications

References 69 publications

Development of a Confinable Gene-Drive System in the Human Disease Vector,Aedes aegypti

Development of a Confinable Gene-Drive System in the Human Disease Vector,Aedes aegypti

Programmable RNA Targeting using CasRx in Flies

Engineered resistance to Zika virus in transgenicAe. aegyptiexpressing a polycistronic cluster of synthetic miRNAs

Contact Info

Product

Resources

About