A homing rescue gene drive with multiplexed gRNAs reaches high frequency in cage populations but generates functional resistance DOI Creative Commons

Jingheng Chen,

Shibo Hou,

Ruobing Feng

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Nov. 29, 2023

Abstract CRISPR homing gene drive is a potent technology with considerable potential for managing populations of medically and agriculturally significant insects. It induces bias in the inheritance allele progeny, rapidly spreading desired genes throughout population. Homing drives operate by Cas9 cleavage followed homology-directed repair, copying to wild-type chromosome. However, resistance alleles formed end-joining repair pose obstacle spread drive. To address this challenge, we created targeting essential but haplosufficient hairy gene. Our strategy involves construct through process, eliminating nonfunctional resistance, which are recessive lethal, while rescuing drive-carrying individuals recoded version . This eliminates more slowly than previous haplolethal genes, it may be easier non-model organisms. rate was moderate, multigenerational cage studies showed quick 96-97% failed reach whole population due formation functional alleles, despite use four gRNAs, that previously successful at preventing resistance. Sequencing these had large deletion must have utilized an alternate start codon. The modest fitness advantage over study, could prevent long-term persistence drive, especially if cargo additional cost. Thus, revised design strategies regions target often necessary avoid such even when using multiplexed gRNAs.

Language: Английский

Next-generation genetic sexing strain establishment in the agricultural pest Ceratitis capitata DOI Creative Commons
Serafima Davydova, Junru Liu, Nikolay P. Kandul

et al.

Scientific Reports, Journal Year: 2023, Volume and Issue: 13(1)

Published: Nov. 14, 2023

Abstract Tephritid fruit fly pests pose an increasing threat to the agricultural industry due their global dispersion and a highly invasive nature. Here we showcase feasibility of early-detection SEPARATOR sex sorting approach through using non-model pest, Ceratitis capitata. This system relies on female-only fluorescent marker expression, accomplished use sex-specific intron highly-conserved transformer gene from C. capitata Anastrepha ludens. The herein characterized strains have 100% desired phenotype outcomes, allowing accurate male–female separation during early development. Overall, describe antibiotic temperature-independent sex-sorting in , which, moving forward, may be implemented other pest species. strategy can facilitate establishment genetic sexing systems with endogenous elements exclusively, wider scale, improve population control strategies like sterile insect technique.

Language: Английский

Citations

9
Allele Sails: launching traits and fates into wild populations with DNA sequence modifiers DOI Creative Commons
Michelle L. Johnson, Bruce A. Hay, Maciej Maselko

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 19, 2024

Population-scale genome editing can be used to alter the composition or fate of wild populations. One approach achieving these aims utilizes a synthetic gene drive element-a multi-gene cassette-to bring about an increase in frequency existing allele. However, use drives is complicated by multiple scientific, regulatory, and social issues associated with transgene persistence flow. Alternatives which transgenes are not driven could potentially avoid some issues. Here we propose population scale using system refer as Allele Sail. An Sail consists editor (the Wind) that introduces DNA sequence edits Sail) at one more sites, resulting progeny viable fertile. The editor, such sequence-specific nuclease, prime- base-editor, inherited Mendelian fashion. Meanwhile, it creates experience arithmetic, Super-Mendelian frequency. We explore this agent-based modeling, identify contexts single, low release brings very high also conditions manipulation sex determination suppression. Current regulatory frameworks often distinguish between transgenics genetically modified organisms (GMOs), their edited non-transgenic non-GMO. In context provides path traits fates populations ways may considered acceptable.

Language: Английский

Citations

0
Population suppression with dominant female-lethal alleles is boosted by homing gene drive DOI Creative Commons
Jinyu Zhu,

Jingheng Chen,

Yiran Liu

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Dec. 5, 2023

Abstract Methods to suppress pest insect populations using genetic constructs and repeated releases of male homozygotes have recently been shown be an attractive alternative older sterile technique based on radiation. Female-specific lethal alleles substantially increased power, but still require large, sustained transgenic releases. Gene drive bias their own inheritance spread throughout populations, potentially allowing population suppression with a single, small-size release. However, drives often suffer from efficiency issues, the most well-studied type, homing drives, tend without limit. In this study, we show that coupling female-specific gene allowed substantial improvement in while retaining self-limiting nature (and thus confinement) allele strategy. Using mosquito model, required sizes for elimination variety scenarios, including different density growth curves, comparisons other systems. Resistance reduced power method, these could overcome by targeting essential also providing rescue. A proof-of-principle demonstration system Drosophila melanogaster was effective both basing its achieving high lethality among females inherit construct absence antibiotic. Overall, our study shows improvements can achieved systems combining them drive.

Language: Английский

Citations

1
Next-generation genetic sexing strain establishment in the agricultural pestCeratitis capitata DOI Open Access
Serafima Davydova, Junru Liu, Nikolay P. Kandul

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Oct. 1, 2023

Abstract Tephritid fruit fly pests pose an increasing threat to the agricultural industry due their global dispersion and a highly invasive nature. Here we showcase feasibility of early-detection SEPARATOR sex sorting approach through using non-model pest, Ceratitis capitata . This system relies on female-only fluorescent marker expression, accomplished use sex-specific intron highly-conserved transformer gene from C. Anastrepha ludens The herein characterized strains have 100% desired phenotype outcomes, allowing accurate male-female separation during early development. Overall, describe antibiotic temperature-independent sex-sorting in , which, moving forward, may be implemented other pest species. strategy can facilitate establishment genetic sexing systems with endogenous elements exclusively, wider scale, improve population control strategies like sterile insect technique.

Language: Английский

Citations

0
A homing rescue gene drive with multiplexed gRNAs reaches high frequency in cage populations but generates functional resistance DOI Creative Commons

Jingheng Chen,

Shibo Hou,

Ruobing Feng

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Nov. 29, 2023

Abstract CRISPR homing gene drive is a potent technology with considerable potential for managing populations of medically and agriculturally significant insects. It induces bias in the inheritance allele progeny, rapidly spreading desired genes throughout population. Homing drives operate by Cas9 cleavage followed homology-directed repair, copying to wild-type chromosome. However, resistance alleles formed end-joining repair pose obstacle spread drive. To address this challenge, we created targeting essential but haplosufficient hairy gene. Our strategy involves construct through process, eliminating nonfunctional resistance, which are recessive lethal, while rescuing drive-carrying individuals recoded version . This eliminates more slowly than previous haplolethal genes, it may be easier non-model organisms. rate was moderate, multigenerational cage studies showed quick 96-97% failed reach whole population due formation functional alleles, despite use four gRNAs, that previously successful at preventing resistance. Sequencing these had large deletion must have utilized an alternate start codon. The modest fitness advantage over study, could prevent long-term persistence drive, especially if cargo additional cost. Thus, revised design strategies regions target often necessary avoid such even when using multiplexed gRNAs.

Language: Английский

Citations

0