Abstract Cas9/gRNA-mediated gene-drive systems have advanced development of genetic technologies for controlling vector-borne pathogen transmission. These include population suppression approaches, analogs insecticidal techniques that reduce the number insect vectors, and modification (replacement/alteration) which interfere with competence to transmit pathogens. Here, we develop a recoded rescue system malaria vector, Anopheles stephensi , relieves load in females caused by integration...

Abstract CRISPR-based gene-drive systems, which copy themselves via gene conversion mediated by the homology-directed repair (HDR) pathway, have potential to revolutionize vector control. However, mutant alleles generated competing non-homologous end-joining (NHEJ) resistant Cas9 cleavage, can interrupt spread of elements. We hypothesized that drives targeting genes essential for viability or reproduction also carrying recoded sequences restore endogenous functionality should benefit from...

Abstract Wolbachia pipientis is an insect endosymbiont known to limit the replication of viruses including dengue and Zika in their primary mosquito vector, Aedes aegypti. being released into populations globally a bid control diseases caused by these viruses. It theorized that Wolbachia’s priming immune system may confer protection against subsequent viral infection. Other hypotheses posit role for competition between host cellular resources. Using A. aegypti cell line infected with , we...

Increasing global temperatures and unpredictable climatic extremes have contributed to the spread of vector-borne diseases. The mosquito Aedes aegypti is main vector multiple arboviruses that negatively impact human health, mostly in low socioeconomic areas world. Co-circulation co-infection these viruses humans been increasingly reported; however, how vectors contribute this alarming trend remains unclear.

Highlights•Antibody microarray to investigate mosquito host cell signaling response Wolbachia•Wolbachia-infected cells show downregulation of the insulin receptor (IR)•Inhibition or silencing IR impairs replication dengue and Zika viruses in vitro•Mosquitoes fed inhibitor have impaired virus replicationSummaryWolbachia-infected mosquitoes are refractory super-infection with arthropod-borne pathogens, but role proteins pathogen-blocking mechanisms remains be elucidated. Here, we use an...

ABSTRACT Sterile alpha motif and histidine-aspartic domain-containing protein 1 (SAMHD1) is a deoxynucleoside triphosphate (dNTP) triphosphohydrolase recently recognized as an antiviral factor that acts by depleting dNTP availability for viral reverse transcriptase (RT). SAMHD1 restriction counteracted the human immunodeficiency virus type 2 (HIV-2) accessory Vpx, which targets proteosomal degradation, resulting in increased of dNTPs consequently enhanced replication. Nucleoside inhibitors...

Globalization and climate change have contributed to the simultaneous increase spread of arboviral diseases. Cocirculation several arboviruses in same geographic region provides an impetus study impacts multiple concurrent infections within individual vector mosquito. Here, we describe coinfection superinfection with Mayaro virus (Togaviridae, Alphavirus) Zika (Flaviviridae, Flavivirus) vertebrate mosquito cells, as well Aedes aegypti adult mosquitoes, understand interaction dynamics these...

ABSTRACT The mosquito Aedes aegypti is a competent vector of multiple pathogens, including dengue, Zika, yellow fever, chikungunya, and Mayaro viruses. Ae. highly invasive currently present in the Americas, Oceania, Asia, Europe, but its distribution pathogens it transmits are expected to change due climate change. Relative humidity (RH) an environmental variable that affects biology can differ between location, habitat, season, with mosquitoes facing significant variation RH during their...

The mosquito vector Aedes aegypti is responsible for transmitting a range of arboviruses including dengue (DENV) and Zika (ZIKV). global reach these viruses increasing due to an expansion the mosquito's geographic urbanization human travel. Vector control remains primary means limiting diseases. Wolbachia pipientis endosymbiotic bacterium insects that has ability block replication pathogens, flaviviruses such as DENV or ZIKV, inside body vector. A strain called wMel currently being released...

Abstract Gene drives are programmable genetic elements that can spread beneficial traits into wild populations to aid in vector-borne pathogen control. Two different have been developed for population modification of mosquito vectors. The Reckh drive (vasa-Cas9) Anopheles stephensi displays efficient allelic conversion through males but generates frequent drive-resistant mutant alleles when passed females. In contrast, the AgNosCd-1 (nos-Cas9) gambiae achieves almost complete both genders....

The core components of CRISPR-based gene drives, Cas9 and guide RNA (gRNA), either can be linked within a self-contained single cassette (full gene-drive, fGD) or provided in two separate elements (split sGD), the latter offering greater control options. We previously engineered split systems that could converted genetically into autonomous full drives. Here, we examine such dual inserted at spo11 locus are recoded to restore function thus organismic fertility. Despite minimal differences...

Despite its ecological flexibility and geographical co-occurrence with human pathogens, little is known about the ability of Anopheles albimanus to transmit arboviruses. To address this gap, we challenged An. females four alphaviruses one flavivirus monitored progression infections. We found species an efficient vector Mayaro virus, O'nyong-nyong Sindbis although latter two do not currently exist in habitat range. was able become infected Chikungunya but virus dissemination rare (indicating...

Transcriptomic profiling has generated extensive lists of genes that respond to viral infection in mosquitoes. These gene contain two types genes; (1) those are responsible for the insect's natural antiviral defense mechanisms, including some known innate immunity genes, and (2) whose change expression may occur simply as a result infection. As genetic modification tools mosquitoes continue improve, opportunities make refractory insects via allelic replacement or delivery small RNAs alter...

Information about genetic engineering (GE) for vector control in the United States is disseminated primarily English, though non-English speakers are equally, and some geographic regions even more affected by such technologies. Non-English-speaking publics should have equal access to information, which especially critical when technology question may impact whole communities. We convened an interdisciplinary workgroup translate previously developed narrated slideshows on gene drive...

ABSTRACT The development of Cas9/gRNA-mediated gene-drive systems has bolstered the advancement genetic technologies for controlling vector-borne pathogen transmission. These include population suppression approaches, analogs insecticidal techniques that reduce number vector insects, and modification (replacement/alteration) which interfere with competence to transmit pathogens. We developed a recoded rescue system in malaria vector, Anopheles stephensi , relieves load females caused by...

Abstract CRISPR-based gene drive systems, which copy themselves based on conversion mediated by the homology directed repair (HDR) pathway, have potential to revolutionize vector control. However, mutant alleles generated competing non-homologous end-joining (NHEJ) pathway that are rendered resistant Cas9 cleavage can interrupt spread of genedrive elements. We hypothesized drives targeting genes essential for viability or reproduction also carrying recoded sequences restore endogenous...

Increasing global temperatures and unpredictable climatic extremes have contributed to the spread of vector-borne diseases. The mosquito

Abstract Gene-drive systems offer an important new avenue for spreading beneficial traits into wild populations. Their core components, Cas9 and guide RNA (gRNA), can either be linked within a single cassette (full gene drive, fGD) or provided in two separate elements (split sGD) wherein the gRNA-bearing element drives presence of independent static source Cas9. We previously designed system engineered to turn split full drives. Here, we provide experimental proof-of-principle such...

Abstract The mosquito Aedes aegypti is a competent vector of multiple pathogens including dengue, Zika, yellow fever, chikungunya, and Mayaro viruses. Ae. highly invasive currently present in the Americas, Oceania, Asia, Europe, but its distribution it transmits are expected to change due climate change. Relative humidity an environmental variable that affects biology can differ between location, habitat, season, with mosquitoes facing significant variation relative during their lifespan....

Abstract Despite its ecological flexibility and geographical co-occurrence with human pathogens, little is known about the ability for Anopheles albimanus to transmit arboviruses. To address this gap, we challenged An. females four alphaviruses one flavivirus monitored progression of infections. We found species an efficient vector Mayaro virus, O’nyong-nyong Sindbis though latter two do not currently exist in habitat range. was able become infected Chikungunya but virus dissemination rare...