Optimized husbandry and targeted gene-editing for the cnidarian Nematostella vectensis.

IF 3.7 2区 生物学 Q1 DEVELOPMENTAL BIOLOGY
Development Pub Date : 2025-01-15 Epub Date: 2025-02-04 DOI:10.1242/dev.204387
João E Carvalho, Maxence Burtin, Olivier Detournay, Aldine R Amiel, Eric Röttinger
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Abstract

Optimized laboratory conditions for research models are crucial for the success of scientific projects. This includes controlling the entire life cycle, having access to all developmental stages and maintaining stable physiological conditions. Reducing the life cycle of a research model can also enhance the access to biological material and speed up genetic tool development. Thus, we optimized the rearing conditions for the sea anemone Nematostella vectensis, a cnidarian research model, to study embryonic and post-metamorphic processes, such as regeneration. We adopted a semi-automated aquaculture system for N. vectensis and developed a dietary protocol optimized for the different life stages. Thereby, we increased spawning efficiencies, juvenile growth and survival rates, and considerably reduced the overall life cycle down to 2 months. To further improve the obtention of CRISPR-Cas9 mutants, we optimized the design of sgRNAs leading to full knockout animals in F0 polyps using a single sgRNA. Finally, we show that NHEJ-mediated transgene insertion is possible in N. vectensis. In summary, our study provides additional resources for the scientific community that uses or plans to use N. vectensis as a research model.

刺胞线虫病的优化饲养和靶向基因编辑。
优化研究模型的实验室条件对科学项目的成功至关重要。这包括控制整个生命周期,进入所有发育阶段,保持稳定的生理条件。缩短研究模型的生命周期还可以增加对生物材料的获取并加快遗传工具的开发。因此,我们优化了海葵线虫(Nematostella vectensis)的饲养条件,以研究海葵的胚胎和后变质过程,如再生。采用了半自动化养殖系统,并针对不同生命阶段制定了优化的饲料方案。因此,我们提高了产卵效率,幼鱼的生长和存活率,并大大缩短了整个生命周期,减少到两个月。为了进一步提高对CRISPR-Cas9突变体的关注,我们优化了sgRNA的设计,从而使用单个sgRNA在F0息肉中实现全KO动物。最后,我们证明了nhej介导的转基因插入是可能的。总而言之,我们的研究为使用或将使用威氏线虫作为研究模型的科学界提供了额外的资源。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Development
Development 生物-发育生物学
CiteScore
6.70
自引率
4.30%
发文量
433
审稿时长
3 months
期刊介绍: Development’s scope covers all aspects of plant and animal development, including stem cell biology and regeneration. The single most important criterion for acceptance in Development is scientific excellence. Research papers (articles and reports) should therefore pose and test a significant hypothesis or address a significant question, and should provide novel perspectives that advance our understanding of development. We also encourage submission of papers that use computational methods or mathematical models to obtain significant new insights into developmental biology topics. Manuscripts that are descriptive in nature will be considered only when they lay important groundwork for a field and/or provide novel resources for understanding developmental processes of broad interest to the community. Development includes a Techniques and Resources section for the publication of new methods, datasets, and other types of resources. Papers describing new techniques should include a proof-of-principle demonstration that the technique is valuable to the developmental biology community; they need not include in-depth follow-up analysis. The technique must be described in sufficient detail to be easily replicated by other investigators. Development will also consider protocol-type papers of exceptional interest to the community. We welcome submission of Resource papers, for example those reporting new databases, systems-level datasets, or genetic resources of major value to the developmental biology community. For all papers, the data or resource described must be made available to the community with minimal restrictions upon publication. To aid navigability, Development has dedicated sections of the journal to stem cells & regeneration and to human development. The criteria for acceptance into these sections is identical to those outlined above. Authors and editors are encouraged to nominate appropriate manuscripts for inclusion in one of these sections.
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