SYNCAS: Efficient CRISPR/Cas9 gene-editing in difficult to transform arthropods

Insect Biochemistry and Molecular Biology, Journal Year: 2024, Volume and Issue: 165, P. 104068 - 104068

Published: Jan. 1, 2024

The genome editing technique CRISPR/Cas9 has led to major advancements in many research fields and this state-of-the-art tool proven its use genetic studies for various arthropods. However, most transformation protocols rely on microinjection of component into embryos, a method which is challenging species. Alternatively, injections can be performed adult females, but efficiencies very low as was shown the two-spotted spider mite, Tetranychus urticae, minute important chelicerate pest crops. In study, we explored different formulations optimize maternal injection protocol T. urticae. We observed strong synergy between branched amphipathic peptide capsules saponins, resulting significant increase knock-out efficiency, exceeding 20%. This formulation, termed SYNCAS, used urticae genes – phytoene desaturase, CYP384A1 Antennapedia also allowed develop co-CRISPR strategy facilitated generation knock-in mutants. addition, SYNCAS successfully applied white white-like western flower thrips, Frankliniella occidentalis. allows routine these species game changer other hard transform

Language: Английский

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Bacterial genome engineering using CRISPR-associated transposases

Nature Protocols, Journal Year: 2024, Volume and Issue: 19(3), P. 752 - 790

Published: Jan. 12, 2024

Language: Английский

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When and how can we predict adaptive responses to climate change?

Evolution Letters, Journal Year: 2023, Volume and Issue: 8(1), P. 172 - 187

Published: Nov. 29, 2023

Predicting if, when, and how populations can adapt to climate change constitutes one of the greatest challenges in science today. Here, we build from contributions special issue on evolutionary adaptation change, a survey its authors, recent literature explore limits opportunities for predicting adaptive responses change. We outline what might be predictable now, future, perhaps never even with our best efforts. More accurate predictions are expected traits characterized by well-understood mapping between genotypes phenotypes experiencing strong, direct selection due A meta-analysis revealed an overall moderate trait heritability evolvability studies performed under future conditions but indicated no significant current conditions, suggesting neither more nor less genetic variation adapting climates. population persistence rescue remains uncertain, especially many species without sufficient ecological data. Still, when polled, authors contributing this were relatively optimistic about ability predict Predictions will improve as expand efforts understand diverse organisms, their ecology, potential. Advancements functional genomic resources, extension non-model union experiments "omics," should also enhance predictions. Although challenging, small advances reduce substantial uncertainties surrounding

Language: Английский

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CRISPR/Cas9-Mediated Editing of BmEcKL1 Gene Sequence Affected Silk Gland Development of Silkworms (Bombyx mori)

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(3), P. 1907 - 1907

Published: Feb. 5, 2024

The silkworm (Bombyx mori) has served humankind through silk protein production. However, traditional sericulture and the industry have encountered considerable bottlenecks must rely on major technological breakthroughs to keep up with current rapid developments. adoption of gene editing technology nevertheless brought new hope industry. long period low efficiency genetic breeding methods obtain high silk-yielding strains hindered development industry; use specifically control expression genes related gland or synthesis is beneficial for obtaining excellent traits. In this study, BmEcKL1 was knocked out in middle (MSGs) posterior (PSGs) glands using CRISPR/Cas9 technology, ΔBmEcKL1-MSG ΔBmEcKL1-PSG improved MSGs PSGs increased production were obtained. This work identifies proves that directly indirectly participates synthesis, providing perspectives investigating mechanisms silkworms, which great significance selecting varieties.

Language: Английский

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The genomics and evolution of inter-sexual mimicry and female-limited polymorphisms in damselflies

Nature Ecology & Evolution, Journal Year: 2023, Volume and Issue: 8(1), P. 83 - 97

Published: Nov. 6, 2023

Sex-limited morphs can provide profound insights into the evolution and genomic architecture of complex phenotypes. Inter-sexual mimicry is one particular type sex-limited polymorphism in which a novel morph resembles opposite sex. While inter-sexual mimics are known both sexes diverse range animals, their evolutionary origin poorly understood. Here, we investigated basis female-limited male common bluetail damselfly. Differential gene expression between has been documented damselflies, but no causal locus previously identified. We found that originated an ancestrally sexually dimorphic lineage association with multiple structural changes, probably driven by transposable element activity. These changes resulted ~900 kb content partly shared close relative, indicating trans-species polymorphism. More recently, third following translocation part male-mimicry sequence position ~3.5 mb apart. evidence balancing selection maintaining mimicry, line previous field population studies. Our results underscore how variants affecting handful potentially regulatory genes morph-specific give rise to phenotypic polymorphisms.

Language: Английский

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RNAi-directed knockdown in the cnidarian fish blood parasite Sphaerospora molnari

Scientific Reports, Journal Year: 2024, Volume and Issue: 14(1)

Published: Feb. 12, 2024

RNA interference (RNAi) is an effective approach to suppress gene expression and monitor regulation. Despite its wide application, use limited in certain taxonomic groups, including cnidarians. Myxozoans are a unique group of cnidarian parasites that diverged from their free-living ancestors about 600 million years ago, with several species causing acute disease farmed wild fish populations. In this pioneering study we successfully applied RNAi blood stages the myxozoan Sphaerospora molnari, combining dsRNA soaking approach, real-time PCR, confocal microscopy, Western blotting. For proof concept, knocked down two unusual actins, one which known play critical role S. molnari cell motility. We observed intracellular uptake after 30 min accumulation all cells typical cell-in-cell structure. actin vitro, transient inhibition for 48 h. disruption cytoskeletal network within primary loss characteristic rotational This workflow could significantly advance functional research Myxozoa, offering new prospects investigating therapeutic targets facilitating drug discovery against economically important parasites.

Language: Английский

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A time course analysis through diapause reveals dynamic temporal patterns of microRNAs associated with endocrine regulation in the butterfly Pieris napi

Authorea (Authorea), Journal Year: 2024, Volume and Issue: unknown

Published: April 20, 2024

Organisms inhabiting highly seasonal environments must cope with a wide range of environmentally induced challenges. Many challenges require extensive physiological modification to survive. In winter, survive extreme cold and limited resources, insects commonly enter diapause, which is an endogenously derived dormant state associated minimized cellular processes low energetic expenditure. Due the high degree complexity involved in substantial regulation required, our understanding primarily derives from transcriptome via messenger RNA expression dynamics. Here we aim advance diapause by investigating microRNA (miRNA) diapausing direct developing pupae butterfly Pieris napi. We identified coordinated patterns miRNA throughout both head abdomen tissues pupae, target identification, found several be enriched for relevant diapause-related processes. also two candidate miRNAs, miR-14-5p miR-2a-3p, that are likely progression through ecdysone synthesis pathway, critical regulator termination. targets phantom, gene has been expressed response ecdysone. Together, these miRNAs match current timing hormonal P. napi provide interesting candidates further explore mechanistic role microRNAs regulation.

Language: Английский

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Multi-omic applications for understanding and enhancing tropical fruit flavour

Plant Molecular Biology, Journal Year: 2024, Volume and Issue: 114(4)

Published: July 8, 2024

Abstract Consumer trends towards nutrient-rich foods are contributing to global increasing demand for tropical fruit. However, commercial cultivars in the breeding pipeline that tailored meet market at risk of possessing reduced fruit flavour qualities. This stems from recurrent prioritised selection superior agronomic traits and not flavour, which may turn reduce consumer satisfaction. There is realisation quality traits, inclusive must be equally selected for; but currently, there limited tools resources available select particularly species. Although sugars, acids, volatile organic compounds known define specific combinations these, result defined preferences, remain unknown many To include preferences selective breeding, it vital determine metabolites underpin them. Then, objective quantitative analysis implemented instead solely relying on human sensory panels. lead development genetic markers through integrated omics approaches target biosynthetic pathways active compounds. In this review, we explore progress able strategically consumer-preferred profiles new

Language: Английский

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The evolutionary consequences of interactions between the epigenome, the genome and the environment

Evolutionary Applications, Journal Year: 2024, Volume and Issue: 17(7)

Published: July 1, 2024

Abstract The epigenome is the suite of interacting chemical marks and molecules that helps to shape patterns development, phenotypic plasticity gene regulation, in part due its responsiveness environmental stimuli. There increasing interest understanding functional evolutionary importance this sensitivity under ecologically realistic conditions. Observations epigenetic variation abounds natural populations have prompted speculation it may facilitate responses rapid perturbations, such as those occurring climate change. A frequent point contention whether variants reflect genetic or are independent it. genome often appear tightly linked interdependent. While many changes genetically determined, converse also true, with DNA sequence influenced by presence marks. Understanding how epigenome, environment interact one another therefore an essential step explaining broader consequences epigenomic variation. Drawing on results from experimental comparative studies carried out diverse plant animal species, we synthesize our current these factors populations, a focus identifying similarities differences between taxonomic groups. We describe main components they vary within taxa. review interacts features determinants, role transposable elements (TEs) integrating environment. And look at recent investigating interactions. Although differentiation nature likely result drift selection stochastic epimutations, there growing evidence significant fraction can be stably inherited could contribute evolution independently

Language: Английский

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MicroRNAs in the developmental toolbox — a comparative approach to understanding their role in regulating insect development

Current Opinion in Insect Science, Journal Year: 2024, Volume and Issue: 66, P. 101256 - 101256

Published: Aug. 28, 2024

Language: Английский

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