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The steroid-inducible pOp6/LhGR gene expression system is fast, sensitive and does NOT cause plant growth defects in rice (Oryza sativa)
bioRxiv - Plant Biology Pub Date : 2021-01-21 , DOI: 10.1101/2021.01.02.425069
Marketa Samalova , Ian Moore

Inducible systems for transgene expression activated by a chemical inducer or an inducer of non-plant origin are desirable tools for both basic plant research and biotechnology. Although, the technology has been widely exploited in model plants, it has not been optimised for use with the major monocotyledonous crop species, namely rice. We have adapted the dexamethasone-inducible pOp6/LhGR system for rice and shown that it is fast, sensitive and tightly regulated, with high levels of induction that remain stable over several generations. Most importantly, we have shown that the system does not cause negative growth defects in vitro or in soil grown plants. Interestingly in the process of testing, we found that another steroid, triamcinolone acetonide, is a more potent inducer in rice than dexamethasone. We present serious considerations for the construct design to avoid undesirable effects caused by the system in plants, leakiness and possible silencing, as well as simple steps how to maximize translation efficiency of a gene of interest. Finally, we compare the performance of the pOp6/LhGR system with other chemically inducible systems tested in rice in terms of the properties of an ideal inducible system.

中文翻译:

类固醇诱导的pOp6 / LhGR基因表达系统快速,灵敏,不会引起水稻中的植物生长缺陷。

由化学诱导物或非植物来源的诱导物激活的用于转基因表达的诱导系统是基础植物研究和生物技术的理想工具。尽管该技术已在模型植物中得到广泛利用,但尚未针对主要单子叶作物(即水稻)进行优化。我们已经对地塞米松诱导的pOp6 / LhGR系统进行了水稻适配,结果表明该系统快速,灵敏且受到严格调控,诱导水平高,可在几代内保持稳定。最重要的是,我们已经证明该系统不会在体外或土壤植物中引起负面的生长缺陷。在测试过程中有趣的是,我们发现另一种类固醇醋酸曲安奈德比地塞米松是水稻中更有效的诱导剂。我们对构建体设计提出了认真的考虑,以避免系统在植物中引起的不良影响,渗漏和可能的沉默,以及如何使目的基因的翻译效率最大化的简单步骤。最后,根据理想诱导系统的特性,我们将pOp6 / LhGR系统与在水稻中测试的其他化学诱导系统的性能进行了比较。
更新日期:2021-01-22
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