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Systematic Tools for Reprogramming Plant Gene Expression in a Simple Model, Marchantia polymorpha.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-04-02 , DOI: 10.1021/acssynbio.9b00511
Susanna Sauret-Güeto 1 , Eftychios Frangedakis 1 , Linda Silvestri 1, 2 , Marius Rebmann 1 , Marta Tomaselli 1 , Kasey Markel 1 , Mihails Delmans 1 , Anthony West 3 , Nicola J Patron 3 , Jim Haseloff 1
Affiliation  

We present the OpenPlant toolkit, a set of interlinked resources and techniques to develop Marchantia as testbed for bioengineering in plants. Marchantia is a liverwort, a simple plant with an open form of development that allows direct visualization of gene expression and dynamics of cellular growth in living tissues. We describe new techniques for simple and efficient axenic propagation and maintenance of Marchantia lines with no requirement for glasshouse facilities. Marchantia plants spontaneously produce clonal propagules within a few weeks of regeneration, and lines can be amplified million-fold in a single generation by induction of the sexual phase of growth, crossing, and harvesting of progeny spores. The plant has a simple morphology and genome with reduced gene redundancy, and the dominant phase of its life cycle is haploid, making genetic analysis easier. We have built robust Loop assembly vector systems for nuclear and chloroplast transformation and genome editing. These have provided the basis for building and testing a modular library of standardized DNA elements with highly desirable properties. We have screened transcriptomic data to identify a range of candidate genes, extracted putative promoter sequences, and tested them in vivo to identify new constitutive promoter elements. The resources have been combined into a toolkit for plant bioengineering that is accessible for laboratories without access to traditional facilities for plant biology research. The toolkit is being made available under the terms of the OpenMTA and will facilitate the establishment of common standards and the use of this simple plant as testbed for synthetic biology.

中文翻译:

用于在简单模型中重编程植物基因表达的系统工具,Marchantia polymorpha。

我们展示了 OpenPlant 工具包,这是一组相互关联的资源和技术,用于将 Marchantia 开发为植物生物工程的试验平台。地香属植物,一种简单的植物,具有开放的发育形式,可以直接可视化活组织中的基因表达和细胞生长动态。我们描述了无需温室设施即可简单有效地无菌繁殖和维护 Marchantia 系的新技术。Marchania 植物在再生后的几周内自发地产生克隆繁殖体,并且通过诱导生长、杂交和后代孢子收获的有性阶段,可以在单代中将品系扩增数百万倍。该植物具有简单的形态和基因组,基因冗余减少,其生命周期的显性阶段是单倍体,使基因分析更容易。我们已经为核和叶绿体转化和基因组编辑构建了强大的 Loop 组装载体系统。这些为构建和测试具有非常理想特性的标准化 DNA 元件的模块化库提供了基础。我们筛选了转录组数据以识别一系列候选基因,提取推定的启动子序列,并在体内测试它们以识别新的组成型启动子元件。这些资源已合并为一个植物生物工程工具包,可供没有传统植物生物学研究设施的实验室使用。该工具包是根据 OpenMTA 的条款提供的,将促进通用标准的建立和使用这种简单的植物作为合成生物学的试验台。我们已经为核和叶绿体转化和基因组编辑构建了强大的 Loop 组装载体系统。这些为构建和测试具有非常理想特性的标准化 DNA 元件的模块化库提供了基础。我们筛选了转录组数据以识别一系列候选基因,提取推定的启动子序列,并在体内测试它们以识别新的组成型启动子元件。这些资源已合并为一个植物生物工程工具包,可供没有传统植物生物学研究设施的实验室使用。该工具包是根据 OpenMTA 的条款提供的,将促进通用标准的建立和使用这种简单的植物作为合成生物学的试验台。我们已经为核和叶绿体转化和基因组编辑构建了强大的 Loop 组装载体系统。这些为构建和测试具有非常理想特性的标准化 DNA 元件的模块化库提供了基础。我们筛选了转录组数据以识别一系列候选基因,提取推定的启动子序列,并在体内测试它们以识别新的组成型启动子元件。这些资源已合并为一个植物生物工程工具包,可供没有传统植物生物学研究设施的实验室使用。该工具包是根据 OpenMTA 的条款提供的,将促进通用标准的建立和使用这种简单的植物作为合成生物学的试验台。
更新日期:2020-04-23
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