当前位置：
X-MOL 学术
›
ACS Synth. Biol.
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Design of Mammalian ON-Riboswitches Based on Tandemly Fused Aptamer and Ribozyme.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2019-12-16 , DOI: 10.1021/acssynbio.9b00371 Kamila Mustafina 1 , Keisuke Fukunaga 1 , Yohei Yokobayashi 1
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2019-12-16 , DOI: 10.1021/acssynbio.9b00371 Kamila Mustafina 1 , Keisuke Fukunaga 1 , Yohei Yokobayashi 1
Affiliation
|
Self-cleaving ribozymes engineered to be activated or inhibited by a small molecule binding to an RNA aptamer inserted within a ribozyme (aptazymes) have proven to be useful for controlling gene expression in living cells. In mammalian cells, an aptazyme embedded in the 5' or 3' untranslated region of an mRNA functions as a synthetic riboswitch to chemically regulate gene expression. However, the variety of aptazyme architectures and the ribozyme scaffolds that have been used for mammalian riboswitches has been limited. In particular, fewer synthetic riboswitches that activate gene expression in response to a small molecule (ON-switches) in mammalian cells have been reported compared to OFF-switches. In this work, we developed mammalian riboswitches that function as guanine-activated ON-switches based on a novel aptazyme architecture in which an aptamer and a ribozyme are fused in tandem. The riboswitch performance was optimized by fine-tuning the stability of a critical stem that controls the ribozyme structure and function, yielding switches with ON/OFF ratios greater than 6.0. Our new aptazyme architecture expands the RNA device toolbox for controlling gene expression in mammalian cells.
中文翻译：
基于串联融合适体和核酶的哺乳动物ON-核糖开关的设计。
经工程改造以被小分子结合至插入核酶内的RNA适体（适配酶）激活或抑制的自裂解核酶已被证明可用于控制活细胞中的基因表达。在哺乳动物细胞中，嵌入mRNA的5'或3'非翻译区的aptazyme发挥合成核糖开关的作用，以化学方式调节基因表达。然而，已经被用于哺乳动物核糖开关的aptazyme结构和核酶支架的多样性受到限制。特别地，已经报道了与OFF-开关相比，响应于哺乳动物细胞中的小分子而激活基因表达的合成核糖开关（ON-开关）更少。在这项工作中，我们基于新型适体结构将适体和核酶串联在一起，开发了可作为鸟嘌呤激活的ON-开关的哺乳动物核糖开关。通过微调控制核酶结构和功能的关键茎的稳定性，优化了核糖开关的性能，从而产生了开关比大于6.0的开关。我们新的aptazyme体系结构扩展了RNA装置工具箱，可用于控制哺乳动物细胞中的基因表达。
更新日期：2019-12-17
中文翻译：
基于串联融合适体和核酶的哺乳动物ON-核糖开关的设计。
经工程改造以被小分子结合至插入核酶内的RNA适体（适配酶）激活或抑制的自裂解核酶已被证明可用于控制活细胞中的基因表达。在哺乳动物细胞中，嵌入mRNA的5'或3'非翻译区的aptazyme发挥合成核糖开关的作用，以化学方式调节基因表达。然而，已经被用于哺乳动物核糖开关的aptazyme结构和核酶支架的多样性受到限制。特别地，已经报道了与OFF-开关相比，响应于哺乳动物细胞中的小分子而激活基因表达的合成核糖开关（ON-开关）更少。在这项工作中，我们基于新型适体结构将适体和核酶串联在一起，开发了可作为鸟嘌呤激活的ON-开关的哺乳动物核糖开关。通过微调控制核酶结构和功能的关键茎的稳定性，优化了核糖开关的性能，从而产生了开关比大于6.0的开关。我们新的aptazyme体系结构扩展了RNA装置工具箱，可用于控制哺乳动物细胞中的基因表达。
京公网安备 11010802027423号