Germline Cas9 promoters with improved performance for homing gene drive

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

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Gene drive-based population suppression in the malaria vector Anopheles stephensi

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: Jan. 24, 2025

Gene drives are alleles that can bias the inheritance of specific traits in target populations for purpose modification or suppression. Here, we construct a homing suppression drive major urban malaria vector Anopheles stephensi targeting female-specific exon doublesex, incorporating two gRNAs and nanos-Cas9 to reduce functional resistance improve female heterozygote fitness. Our results show was recessive sterile both females males, with various intersex phenotypes homozygotes. Both male heterozygotes only moderate conversion, indicating nanos promoter has lower activity A. than gambiae. By amplicon sequencing, detect very low level allele formation. Combination vasa-Cas9 line boosts conversion rate 100%, suggesting use similar systems population continuous release strategy SIT fsRIDL techniques. This study contributes valuable insights development more efficient environmentally friendly pest control tools aimed at disrupting disease transmission. their spread through populations. constructed efficiency 2-gRNA mosquito doublesex. With vasa-Cas9, high self-limiting achieved.

Language: Английский

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Adversarial interspecies relationships facilitate population suppression by gene drive in spatially explicit models

Ecology Letters, Journal Year: 2023, Volume and Issue: 26(7), P. 1174 - 1185

Published: May 10, 2023

Suppression gene drives bias their inheritance to spread through a population, potentially eliminating it when they reach high frequency. CRISPR homing suppression have already seen success in the laboratory, but several models predict that may be elusive population with realistic spatial structure due extinction-recolonization cycles. Here, we extend our continuous space framework include two competing species or predator-prey pairs. We find both general and mosquito-specific models, predators can facilitate drive-based suppression, albeit at cost of an increased rate drive loss outcomes. These results are robust mosquito seasonal fluctuations. Our study illustrates difficulty predicting outcomes complex ecosystems. However, promising for prospects less powerful successfully eliminate target other pest populations.

Language: Английский

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Population suppression by release of insects carrying a dominant sterile homing gene drive targeting doublesex in Drosophila

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

Published: Sept. 14, 2024

Language: Английский

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A population modification gene drive targeting both Saglin and Lipophorin impairs Plasmodium transmission in Anopheles mosquitoes

eLife, Journal Year: 2023, Volume and Issue: 12

Published: Dec. 5, 2023

Lipophorin is an essential, highly expressed lipid transport protein that secreted and circulates in insect hemolymph. We hijacked the Anopheles coluzzii gene to make it co-express a single-chain version of antibody 2A10, which binds sporozoites malaria parasite Plasmodium falciparum . The resulting transgenic mosquitoes show markedly decreased ability transmit berghei expressing P. circumsporozoite mice. To force spread this antimalarial transgene mosquito population, we designed tested several CRISPR/Cas9-based drives. One these installed in, disrupts, pro-parasitic Saglin also cleaves wild-type Lipophorin, causing anti-malarial modified replace wild type hitch-hike together with drive. Although generating drive-resistant alleles showing instability its gRNA-encoding multiplex array, -based drive reached high levels caged populations efficiently promoted simultaneous Lipophorin::Sc2A10 allele. This combination expected decrease transmission via two different mechanisms. work contributes design novel strategies transgenes mosquitoes, illustrates some unexpected outcomes encountered when establishing population modification

Language: Английский

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Population suppression with dominant female-lethal alleles is boosted by homing gene drive

BMC Biology, Journal Year: 2024, Volume and Issue: 22(1)

Published: Sept. 11, 2024

Language: Английский

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Performance characteristics allow for confinement of a CRISPR toxin–antidote gene drive for population suppression in a reaction–diffusion model

Proceedings of the Royal Society B Biological Sciences, Journal Year: 2024, Volume and Issue: 291(2025)

Published: June 1, 2024

Gene drive alleles that can bias their own inheritance could engineer populations for control of disease vectors, invasive species and agricultural pests. There are successful examples suppression drives confined modification drives, but developing has proven more difficult. However, CRISPR-based toxin–antidote dominant embryo (TADE) may fill this niche. It works by targeting disrupting a haplolethal target gene in the germline with its gRNAs while rescuing target. also disrupts female fertility driving insertion or additional gRNAs. Here, we used reaction–diffusion model to assess performance continuous space, where outcomes be substantially different from those panmictic populations. We measured wave speed found moderate fitness costs disruption early maternally deposited nuclease eliminate drive’s ability form advance. assessed required release size, finally investigated migration corridor scenarios. is often possible suppress one population then persist without invading second population, potentially desirable outcome. Thus, even imperfect variants TADE excellent candidates suppression.

Language: Английский

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Simulations Reveal High Efficiency and Confinement of a Population Suppression CRISPR Toxin-Antidote Gene Drive

ACS Synthetic Biology, Journal Year: 2023, Volume and Issue: 12(3), P. 809 - 819

Published: Feb. 24, 2023

Though engineered gene drives hold great promise for spreading through and suppressing populations of disease vectors or invasive species, complications such as resistance alleles spatial population structure can prevent their success. Additionally, most forms suppression drives, homing driving Y chromosomes, will generally spread uncontrollably between with even small levels migration. The previously proposed CRISPR-based toxin-antidote system called dominant embryo (TADE) drive could potentially address the issues confinement resistance. However, it is a relatively weak form compared to which might make particularly vulnerable structure. In this study, we investigate TADE using individual-based simulations in continuous landscape. We find that actually more confined than simple models without space, its efficient low cleavage rate embryos from maternally deposited Cas9. Furthermore, performed well space scenarios if initial release requirements were met, timely manner being severely affected by chasing, phenomenon wild-type individuals avoid recolonizing empty areas. At higher cut rates, loses ability spread, but single, widespread often still induce rapid collapse. Thus, be successfully constructed, they may play an important role control species when stringent target desired.

Language: Английский

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New germline Cas9 promoters show improved performance for homing gene drive

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

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Gene drive-based population suppression in the malaria vectorAnopheles stephensi

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

Published: May 24, 2024

Abstract Gene drives are alleles that can bias the inheritance of specific traits in target populations for purpose modification or suppression. Here, we constructed a homing suppression drive major urban malaria vector Anopheles stephensi targeting female-specific exon doublesex , incorporating two gRNAs and nanos- Cas9 promoter to reduce functional resistance improve female heterozygote fitness. Our result showed was recessive sterile both females males, with various intersex phenotypes homozygotes. Both male heterozygotes only moderate conversion, indicating nos has lower activity A. than gambiae . By amplicon sequencing, detected very low level allele formation. Combination vasa -Cas9 line demonstrated boost conversion rate 100%, suggesting use similar systems population continuous release strategy SIT fRIDL techniques. This study contributes valuable insights development more efficient environmentally friendly pest control tools aimed at disrupting disease transmission.

Language: Английский

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