Boosting riboswitch efficiency by RNA amplification.
Boosting riboswitch efficiency by RNA amplification.
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DOI：
10.1093/nar/gkv165
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发表时间：
2015-05-26
影响因子：
14.9
通讯作者：
Bock R
中科院分区：
文献类型：
--
作者：
Emadpour M;Karcher D;Bock R
核糖开关是 RNA 传感器，可响应小分子的结合来调节基因表达。尽管它们在概念上代表简单的开/关开关，因此对生物技术和未来的合成生物学应用具有广阔的前景，但在添加或去除配体后天然核糖开关对基因表达的诱导通常只是中等程度，因此可达到的表达水平不是很高。在这里，我们设计了一种基于 RNA 扩增的系统，可以极大地提高核糖开关的效率。我们已经在生物系统中成功实施了该方法，目前尚无有效的诱导（转基因）基因表达的内源工具：高等植物的叶绿体。我们进一步表明，叶绿体基因组的组成型表达对植物有害的HIV抗原可以在RNA扩增增强核糖开关（RAmpER）的控制下诱导表达，而不会引起突变表型，证明了该方法生产对宿主细胞有毒的蛋白质和代谢物的潜力。
Riboswitches are RNA sensors that regulate gene expression in response to binding of small molecules. Although they conceptually represent simple on/off switches and, therefore, hold great promise for biotechnology and future synthetic biology applications, the induction of gene expression by natural riboswitches after ligand addition or removal is often only moderate and, consequently, the achievable expression levels are not very high. Here, we have designed an RNA amplification-based system that strongly improves the efficiency of riboswitches. We have successfully implemented the method in a biological system for which currently no efficient endogenous tools for inducible (trans)gene expression are available: the chloroplasts of higher plants. We further show that an HIV antigen whose constitutive expression from the chloroplast genome is deleterious to the plant can be inducibly expressed under the control of the RNA amplification-enhanced riboswitch (RAmpER) without causing a mutant phenotype, demonstrating the potential of the method for the production of proteins and metabolites that are toxic to the host cell.
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影响因子：
14.9
作者：
Neupert, Juliane;Karcher, Daniel;Bock, Ralph
通讯作者：
Bock, Ralph
影响因子：
4.9
作者：
Lössl, A;Bohmert, K;Koop, HU
通讯作者：
Koop, HU
DOI：
10.1038/nbt0495-362
发表时间：
1995-04-01
期刊：
BIO-TECHNOLOGY
影响因子：
--
作者：
MCBRIDE, KE;SVAB, Z;MALIGA, P
通讯作者：
MALIGA, P
影响因子：
20.5
作者：
Daniell, Henry;Singh, Nameirakpam D.;Mason, Hugh;Streatfield, Stephen J.
通讯作者：
Streatfield, Stephen J.
影响因子：
16.8
作者：
Mandal, M;Breaker, RR
通讯作者：
Breaker, RR