Synthetic promoter designs enabled by a comprehensive analysis of plant core promoters.
Synthetic promoter designs enabled by a comprehensive analysis of plant core promoters.
复制标题
通过对植物核心启动子的全面分析，合成启动子设计成为可能。
DOI：
10.1038/s41477-021-00932-y
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发表时间：
2021-06
期刊：
影响因子：
18
通讯作者：
Queitsch, Christine
中科院分区：
文献类型：
--
作者：
Jores, Tobias;Tonnies, Jackson;Wrightsman, Travis;Buckler, Edward S.;Cuperus, Josh T.;Fields, Stanley;Queitsch, Christine
植物基因表达的靶向工程为确保粮食安全和在植物中生产生物药物带来了巨大的希望。然而，这一工程需要对顺式调控元件的全面了解，以精确控制内源或引入的基因。为了获得这一知识，我们使用了大规模平行报告试验来测量来自拟南芥、玉米和高粱的几乎完整的启动子集的活性。我们证明了核心启动子元件-特别是TATA盒-以及启动子GC含量和启动子-近端转录因子结合位点影响启动子强度。通过在双子叶烟草的叶片和单子叶玉米的原生质体两个检测系统中进行实验，我们检测了GC含量和转录因子对启动子强度的贡献在物种特定的差异。利用这些观察，我们建立了计算模型来预测两个测试系统中的启动子强度，使我们能够设计出活性与病毒35S最小启动子相当的高活性启动子。我们的结果为优化天然启动子元件和产生具有理想特性的合成启动子元件建立了一种有前景的实验方法。
Targeted engineering of plant gene expression holds great promise for ensuring food security and for producing biopharmaceuticals in plants. However, this engineering requires thorough knowledge of cis-regulatory elements to precisely control either endogenous or introduced genes. To generate this knowledge, we used a massively parallel reporter assay to measure the activity of nearly complete sets of promoters from Arabidopsis, maize and sorghum. We demonstrate that core promoter elements—notably the TATA box—as well as promoter GC content and promoter-proximal transcription factor binding sites influence promoter strength. By performing the experiments in two assay systems, leaves of the dicot tobacco and protoplasts of the monocot maize, we detect species-specific differences in the contributions of GC content and transcription factors to promoter strength. Using these observations, we built computational models to predict promoter strength in both assay systems, allowing us to design highly active promoters comparable in activity to the viral 35S minimal promoter. Our results establish a promising experimental approach to optimize native promoter elements and generate synthetic ones with desirable features.
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