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

Germline Cas9 promoters with improved performance for homing gene drive DOI Creative Commons
Jie Du, W.-C. Chen,

Xihua Jia

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: May 29, 2024

Abstract Gene drive systems could be a viable strategy to prevent pathogen transmission or suppress vector populations by propagating alleles with super-Mendelian inheritance. CRISPR-based homing gene drives convert wild type into in heterozygotes Cas9 and gRNA. It is thus desirable identify promoters that yield high conversion rates, minimize the formation rate of resistance both germline early embryo, limit somatic expression. In Drosophila , nanos promoter avoids leaky expression, but at cost embryo from maternally deposited Cas9. To improve efficiency, we test eleven melanogaster promoters. Some achieve higher efficiency minimal resistance, none completely avoid However, such expression often does not carry detectable fitness costs for rescue targeting haplolethal gene, suggesting conversion. Supporting 4-gRNA suppression drive, one leads low equilibrium frequency due other outperforms resulting successful cage population. Overall, these hold advantages species may possess valuable homologs organisms.

Language: Английский

Citations

10
Assessment of drive efficiency and resistance allele formation of a homing gene drive in the mosquito Aedes aegypti DOI

Xiaozhen Yang,

Xuejiao Xu,

Yixian Chen

et al.

Journal of Pest Science, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 8, 2025

Language: Английский

Citations

1
Manipulating the Destiny of Wild Populations Using CRISPR DOI Creative Commons
Robyn Raban, John M. Marshall, Bruce A. Hay

et al.

Annual Review of Genetics, Journal Year: 2023, Volume and Issue: 57(1), P. 361 - 390

Published: Sept. 19, 2023

Genetic biocontrol aims to suppress or modify populations of species protect public health, agriculture, and biodiversity. Advancements in genome engineering technologies have fueled a surge research this field, with one gene editing technology, CRISPR, leading the charge. This review focuses on current state CRISPR for genetic pests highlights progress ongoing challenges using these approaches.

Language: Английский

Citations

11
Engineering drive–selection balance for localized population suppression with neutral dynamics DOI Creative Commons
Katie Willis, Austin Burt

Proceedings of the National Academy of Sciences, Journal Year: 2025, Volume and Issue: 122(6)

Published: Feb. 4, 2025

While the release of sterile males has been highly successful in suppressing some pest populations, it is impractical for many species due to disappearing after a single generation, necessitating large, repeated releases maintain sufficient impact. Synthetic gene drives promise more efficient approaches since they can increase frequency from rare, yet this also allows them spread across landscape, which may not always be desired. Between these two extremes are selectively neutral genetic constructs persist at released, offering potential suppression that remains localized. One way achieve would have perfect balance, all construct frequencies, between drive increasing and selection decreasing it. Here, we describe closely approximate balance using toxin–antidote causes recessive lethality or sterility, encodes genomic editor makes dominant lethal edits genome, provides protection against action consequences editing. Computer modeling shows design 100-fold than males, 1,000-fold when released alongside booster. We designs CRISPR-based molecular construction, including options avoid recoded genes as antidotes.

Language: Английский

Citations

0
Genome Editing in Hymenoptera DOI Creative Commons
Hamish A. Salvesen, Peter K. Dearden

Insect Biochemistry and Molecular Biology, Journal Year: 2025, Volume and Issue: 180, P. 104300 - 104300

Published: March 11, 2025

The application of genome editing tools in Hymenoptera has transformative potential for functional genetics and understanding their unique biology. comprise one the most diverse Orders animals, development methods efficiently creating precise modifications could have applications conservation, pest management agriculture. To date, sex determination, DNA methylation, taste smell sensory systems as well phenotypic markers been selected gene investigations. From these data, insights into eusociality, nature haplodiploidy complex communication that possess provided an evolutionary history led them to become so successful. Insights from analyses supported by ever-improving suite CRIPSR further expansion will allow more specific biological hypotheses be tested beyond lab. Looking ahead, Hymenopteran modifying biocontrol agents agricultural pests use managing invasive species through technologies such drives. This review provides accessibility information regarding status editing, intending support considered CRISPR novel innovation refinement which it already achieved.

Language: Английский

Citations

0
Genetic engineering for SIT application: a fruit fly‐focused review DOI
Serafima Davydova, Danheng Yu, Angela Meccariello

et al.

Insect Science, Journal Year: 2025, Volume and Issue: unknown

Published: April 7, 2025

Abstract Sterile insect technique (SIT) has become a key component of efficient pest control. Fruit fly pests from the Drosophilidae and Tephritidae families pose substantial overwhelmingly increasing threat to agricultural industry, aggravated by climate change globalization among other contributors. In this review, we discuss advances in genetic engineering aimed improve SIT‐mediated fruit This includes SIT enhancement strategies such as novel sexing strain female lethality approaches. Self‐pervasive X‐shredding X‐poisoning sex distorters, alongside gene drive varieties are also reviewed. The self‐limiting precision‐guided SIT, which aims tackle removal male fertility via CRISPR/Cas9, is additionally introduced. By using examples existing tools interest, well model species, illustrate that population control intensity may be modulated depending on strategy selection.

Language: Английский

Citations

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

Jingheng Chen,

Ruobing Feng

et al.

Journal of genetics and genomics/Journal of Genetics and Genomics, Journal Year: 2024, Volume and Issue: 51(8), P. 836 - 843

Published: April 8, 2024

Language: Английский

Citations

3
Engineering drive-selection balance for localised population suppression with neutral dynamics DOI Creative Commons
Katie Willis, Austin Burt

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

Published: May 21, 2024

Abstract Whilst the release of sterile males has been highly successful in suppressing some pest populations, it is impractical for other species due to disappearing after a single generation, necessitating large, repeated releases maintain sufficient impact. Synthetic gene drives promise more efficient approaches since they can increase frequency from rare, yet this also allows them spread across landscape, which may not always be desired. Between these two extremes are selectively neutral genetic constructs persist at released, offering potential suppression that remains localised. One way achieve would have perfect balance, all construct frequencies, between drive increasing and selection decreasing it. Here we describe create balance involving toxin-antidote causes recessive lethality, encodes genomic editor makes dominant lethal edits genome, provides protection against action or consequences editing. Computer modelling shows design 100-fold than males, 1000-fold when released alongside booster. We designs CRISPR-based molecular construction, including options avoid using recoded genes as antidotes.

Language: Английский

Citations

2
New germline Cas9 promoters show improved performance for homing gene drive DOI Creative Commons
Jie Du,

Weizhe Chen,

Xihua Jia

et al.

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

Published: July 16, 2023

Abstract Gene drive systems could be a viable strategy to prevent pathogen transmission or suppress vector populations by propagating alleles with super-Mendelian inheritance. CRISPR-based homing gene drives, perhaps the most powerful strategy, convert wild type into in heterozygotes help of Cas9 and gRNA. However, achieving successful outcomes these drives often requires high performance. Specifically, it is desirable identify promoters that yield conversion rates, minimize formation rate resistance both germline early embryo, limit somatic expression. Thus far, high-performance have only been discovered Anopheles species. In Drosophila , nanos promoter avoids leaky expression, but at cost embryo from maternally deposited Cas9. To improve efficiency, we tested eleven melanogaster several configurations. Some new achieved higher efficiency minimal resistance, none completely avoid expression like . such did not carry detectable fitness costs when promoter-Cas9 elements supported rescue targeting haplolethal gene, suggesting conversion. Based on our findings, selected two lines for cage experiments 4-gRNA suppression drive. While one exhibited substantial effects, leading low equilibrium frequency, other outperformed resulting population. Overall, novel hold potential advantages species may also possess valuable homologs organisms.

Language: Английский

Citations

6
Advancements and Future Prospects of CRISPR-Cas-Based Population Replacement Strategies in Insect Pest Management DOI Creative Commons
Yu Zhao, L.-L. Li,

Liangzi Wei

et al.

Insects, Journal Year: 2024, Volume and Issue: 15(9), P. 653 - 653

Published: Aug. 30, 2024

Population replacement refers to the process by which a wild-type population of insect pests is replaced possessing modified traits or abilities. Effective necessitates gene drive system capable spreading desired genes within natural populations, operating under principles akin super-Mendelian inheritance. Consequently, releasing small number genetically edited insects could potentially achieve control objectives. Currently, several approaches are exploration, including newly adapted CRISPR-Cas genome editing system. Multiple studies investigating methods engineer that incapable causing crop damage transmitting vector-borne diseases, with notable successful examples documented. This review summarizes recent advancements in realm and provides insights into research methodologies, testing protocols, implementation strategies for techniques. The also discusses emerging trends prospects establishing genetic tools pest management.

Language: Английский

Citations

1