A5058249892- CRISPR and Genetic Engineering
- Insect symbiosis and bacterial influences
- Mosquito-borne diseases and control
- Insect Resistance and Genetics
- Evolution and Genetic Dynamics
- RNA and protein synthesis mechanisms
- Complex Systems and Time Series Analysis
- Animal Genetics and Reproduction
- Chromosomal and Genetic Variations
- Plant Taxonomy and Phylogenetics
- Time Series Analysis and Forecasting
- Malaria Research and Control
- Reading and Literacy Development
- Genetics, Bioinformatics, and Biomedical Research
- Insect behavior and control techniques
- Opinion Dynamics and Social Influence
- Educational and Psychological Assessments
- Insect and Arachnid Ecology and Behavior
- Diptera species taxonomy and behavior
- Complex Network Analysis Techniques
- Rice Cultivation and Yield Improvement
- COVID-19 epidemiological studies
- Intellectual Property Law
- Advanced biosensing and bioanalysis techniques
- Copyright and Intellectual Property
University of California, Berkeley
2018-2023
Berkeley College
2019-2022
The Ohio State University Wexner Medical Center
2015-2016
The Ohio State University
2015
International Rice Research Institute
2008
Aedes aegypti is the principal mosquito vector for many arboviruses that increasingly infect millions of people every year. With an escalating burden infections and relative failure traditional control methods, development innovative measures has become paramount importance. The use gene drives sparked significant enthusiasm genetic mosquitoes; however, no such system been developed in Ae. aegypti. To fill this void, here we develop several CRISPR-based split vector. cleavage rates up to...
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 The mosquito Aedes aegypti is the principal vector for arboviruses including dengue/yellow fever, chikungunya, and Zika virus, infecting hundreds of millions people annually. Unfortunately, traditional control methodologies are insufficient, so innovative methods needed. To complement existing measures, here we develop a molecular genetic system termed precision-guided sterile insect technique (pgSIT) in . PgSIT uses simple CRISPR-based approach to generate flightless females males...
Small laboratory cage trials of non-drive and gene-drive strains the Asian malaria vector mosquito, Anopheles stephensi, were used to investigate release ratios other strain properties for their impact on transgene spread during simulated population modification. We evaluated effects transgenes survival, male contributions next-generation populations, female reproductive success accumulation gene drive-resistant genomic target sites resulting from nonhomologous end-joining (NHEJ) mutagenesis...
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 CRISPR-based gene drives can spread through wild populations by biasing their own transmission above the 50% value predicted Mendelian inheritance. These technologies offer population-engineering solutions for combating vector-borne diseases, managing crop pests, and supporting ecosystem conservation efforts. Current raise safety concerns unintended propagation. Herein, we address such splitting drive components, Cas9 gRNAs, into separate alleles to form a trans-complementing...
Abstract A recurring target-site mutation identified in various pests and disease vectors alters the voltage gated sodium channel ( vgsc ) gene (often referred to as knockdown resistance or kdr confer commonly used insecticides, pyrethroids DDT. The ubiquity of mutations poses a major global threat continued use insecticides means for vector control. In this study, we generate common isogenic laboratory Drosophila strains using CRISPR/Cas9 editing. We identify differential sensitivities...
Abstract Malaria, dengue, Zika and other mosquito‐borne diseases continue to pose a major global health burden through much of the world, despite widespread distribution insecticide‐based tools antimalarial drugs. The advent CRISPR/Cas9‐based gene editing its demonstrated ability streamline development drive systems has reignited interest in application this technology control mosquitoes they transmit. versatility enabled wide range architectures be realized, creating need for their...
Homing-based gene drives, engineered using CRISPR/Cas9, have been proposed to spread desirable genes throughout populations. However, invasion of such drives can be hindered by the accumulation resistant alleles. To limit this obstacle, we engineer a confinable population modification home-and-rescue (HomeR) drive in Drosophila targeting an essential gene. In our experiments, alleles that disrupt target function were recessive lethal and therefore disadvantaged. We demonstrate HomeR achieve...
Interest in gene drive technology has continued to grow as promising new systems have been developed the lab and discussions are moving towards implementing field trials. The prospect of trials requires models that incorporate a significant degree ecological detail, including parameters change over time response environmental data such temperature rainfall, leading seasonal patterns mosquito population density. Epidemiological outcomes also growing importance, as: i) suitability construct...
Abstract Background The discovery of CRISPR-based gene editing and its application to homing-based drive systems has been greeted with excitement, for potential control mosquito-borne diseases on a wide scale, concern, the invasiveness irreversibility release. Gene that display threshold-dependent behavior could potentially be used during trial phase this technology, or when localized is otherwise desired, as simple models predict them spread into partially isolated populations in...
CRISPR-based genetic engineering tools aimed to bias sex ratios, or drive effector genes into animal populations, often integrate the transgenes autosomal chromosomes. However, in species with heterogametic chromsomes (e.g. XY, ZW), linkage of endonucleases could be beneficial expression a sex-specific manner produce sexing systems, ratio distorters, even gene drives, for example. To explore this possibility, here we develop transgenic line Drosophila melanogaster expressing Cas9 from Y...
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...
Abstract Fluorophore labeling of proteins while preserving native functions is essential for bulk Förster resonance energy transfer (FRET) interaction and single molecule imaging analysis. Here we describe a versatile, efficient, specific, irreversible, gentle low-cost method with fluorophores that appears substantially more robust than similar but chemically distinct procedure. The employs the controlled enzymatic conversion central Cys to reactive formylglycine (fGly) aldehyde within six...
Abstract Malaria, dengue, Zika, and other mosquito-borne diseases continue to pose a major global health burden through much of the world, despite widespread distribution insecticide-based tools antimalarial drugs. The advent CRISPR/Cas9-based gene editing its demonstrated ability streamline development drive systems has reignited interest in application this technology control mosquitoes they transmit. versatility also enabled wide range architectures be realized, creating need for their...
Abstract 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 infections relative failure traditional control methods, development innovative measures has become paramount importance. The use gene drives recently sparked significant enthusiasm genetic populations, however no such system been developed in Ae. . To fill this void demonstrate...
Abstract Homing based gene drives, engineered using CRISPR/Cas9, have been proposed to spread desirable genes into target populations. However, of such drives can be hindered by the accumulation resistance alleles. To overcome this significant obstacle, we engineer an inherently confinable population modification Home -and- R escue (HomeR) drive in Drosophila melanogaster that, creative design, limits We demonstrate that HomeR achieve nearly ∼100% transmission enabling it and persist at...
Close-kin mark-recapture (CKMR) methods have recently been used to infer demographic parameters such as census population size and survival for fish of interest fisheries conservation. These advantages over traditional the mark is genetic, removing need physical marking recapturing that may interfere with parameter estimation. For mosquitoes, spatial distribution close-kin pairs has estimate mean dispersal distance, relevance vector-borne disease transmission novel biocontrol strategies....
Abstract The mosquito Aedes aegypti is the principal vector for arboviruses including dengue/yellow fever, chikungunya, and zika, infecting hundreds of millions people annually. Unfortunately, traditional control methodologies are insufficient, so innovative methods needed. To complement existing measures, here we develop a molecular genetic system termed precision guided sterile insect technique (pgSIT) in . PgSIT uses simple CRISPR-based approach to generate males that deployable at any...
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 drives spread through populations bypassing the dictates of Mendelian genetics, offering a population-engineering tool for tackling vector-borne diseases, managing crop pests, and helping island conservation efforts; unfortunately, current technologies raise safety concerns unintended propagation. Herein, we address this by splitting two drive components, Cas9 gRNAs, into separate alleles to form novel trans-complementing split–gene-drive (tGD) demonstrate its...
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...
Abstract Interest in gene drive technology has continued to grow as promising new systems have been developed the lab and discussions are moving towards implementing field trials. The prospect of trials requires models that incorporate a significant degree ecological detail, including parameters change over time response environmental data such temperature rainfall, leading seasonal patterns mosquito population density. Epidemiological outcomes also growing importance, as: i) suitability...