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Autoinhibited transient, gated, and cascaded dynamic transcription of RNAs
Science Advances ( IF 13.6 ) Pub Date : 2022-08-17 , DOI: 10.1126/sciadv.abq5947
Zhenzhen Li 1 , Jianbang Wang 1 , Itamar Willner 1
Affiliation  

Following transient spatiotemporal misregulation of gene expression programs by native transcription machineries, we introduce a versatile biomimetic concept to design transient dynamic transcription machineries, revealing gated and cascaded temporal transcription of RNAs. The concept is based on the engineering of the reaction module consisting of malachite green (MG) and/or DFHBI {(5Z)-5-[(3,5-difluoro-4-hydroxyphenyl)methylene]-3,5-dihydro-2,3-dimethyl-4 H -imidazol-4-one} DNA scaffolds, T7 RNA polymerase (RNAP) aptamer transcription scaffold, and the inhibited T7 RNAP-aptamer complex. In the presence of the counter RNAP aptamer strand and ribonucleoside triphosphates, the triggered activation of the three transcription scaffolds are activated, leading to the transcription of the MG and/or DFHBI RNA aptamer and to the transcription of the RNAP aptamer acting as an autoinhibitor that leads to the transient temporal, dissipative, and blockage of all transcription. By appropriate design of the transcription scaffolds and the inhibitors/coupler, transient gated and cascaded transcription processes are demonstrated, and a bimodal transcription module synthesizing a transient operating ribozyme is introduced.

中文翻译:

自动抑制 RNA 的瞬时、门控和级联动态转录

在原生转录机器对基因表达程序的瞬时时空错误调节之后,我们引入了一种通用的仿生概念来设计瞬时动态转录机器,揭示了 RNA 的门控和级联时间转录。该概念基于由孔雀石绿 (MG) 和/或 DFHBI {(5Z)-5-[(3,5-difluoro-4-hydroxyphenyl)methylene]-3,5-dihydro- 2,3-二甲基-4H-imidazol-4-one} DNA 支架、T7 RNA 聚合酶 (RNAP) 适体转录支架和受抑制的 T7 RNAP-适体复合物。在反 RNAP 适体链和核糖核苷三磷酸存在的情况下,三个转录支架的触发激活被激活,导致 MG 和/或 DFHBI RNA 适体的转录和作为自身抑制剂的 RNAP 适体的转录导致所有转录的瞬时、耗散和阻塞。通过适当设计转录支架和抑制剂/偶联剂,证明了瞬时门控和级联转录过程,并引入了合成瞬时操作核酶的双峰转录模块。
更新日期:2022-08-17
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