Tunable Riboregulator Switches for Post-transcriptional Control of Gene Expression.
Tunable Riboregulator Switches for Post-transcriptional Control of Gene Expression.
复制标题
DOI：
10.1021/acssynbio.5b00041
复制
发表时间：
2015-12-18
影响因子：
4.7
通讯作者：
Unkefer CJ
中科院分区：
文献类型：
--
作者：
Krishnamurthy M;Hennelly SP;Dale T;Starkenburg SR;Martí-Arbona R;Fox DT;Twary SN;Sanbonmatsu KY;Unkefer CJ
由于代谢反应的产量与酶的浓度(d[cp]/dt~kcat[ce])成正比，到目前为止，通过特定代谢步骤改变通量的最直接方法是增加或减少酶催化剂的浓度。目前的基因工程表达策略仅限于通过将特定基因置于更强大的转录启动子的控制下来增加该基因的表达，或者使用几种策略中的一种来敲除或敲除浪费的基因。为了微调酶浓度的表达，我们开发了能够独立控制工程代谢途径中每种酶浓度的核糖体调节剂。用一种独特的方法来设计和优化核转录调控因子的活性，这是一种合理设计和定向进化的核转录调控因子集合。核调节器的整体架构采用Watson-Crick碱基配对稳定性设计。事实上，已经使用这种方法设计了许多具有明确性能特征的可调小调节RNA。然后，将使用最先进的结构建模和RNA的分子模拟来优化该架构。接下来将在每个核转录调节因子的一部分上利用定向进化。通过这种方式，一组核调节因子(每个基因一组)将同时进化，以优化一组酶的浓度，使通量最大化。
Because the output of a metabolic reaction is directly proportional to the enzyme concentration (d[cp]/dt ~ kcat[ce]), by far the most straightforward approach to altering the flux through a particular metabolic step is to increase or decrease the concentration of the enzyme catalyst. Current strategies for engineering gene expression are limited to increasing the expression of a particular gene by placing it under the control of a stronger transcriptional promoter or using one of several strategies to knock down or knock out a wasteful gene. To finely tune expression of enzyme concentrations, we have developed riboregulators that has the potential to independently control the concentration of each enzyme in an engineered metabolic pathway. A unique approach to rationale design and directed evolution of the ensemble of riboregulators was used to design and optimize the activity of the riboregulators. The overall architecture of the riboregulators is designed using Watson-Crick base pairing stability. In fact, many tunable small regulatory RNAs with well-defined performance characteristics have been designed using this method. The architecture will then be refined using state-of-the-art structural modeling and molecular simulation of RNA. Directed evolution will next be utilized on a portion of each riboregulator. In this manner, a set of riboregulators (one for each gene) will be evolved simultaneously to optimize the set of enzyme concentrations to maximize flux.
登录
查看更多内容
影响因子：
64.5
作者：
Pardee K;Green AA;Ferrante T;Cameron DE;DaleyKeyser A;Yin P;Collins JJ
通讯作者：
Collins JJ
影响因子：
8.4
作者：
Carter, Karen K.;Valdes, James J.;Bentley, William E.
通讯作者：
Bentley, William E.
影响因子：
14.8
作者：
Nielsen, Jens;Fussenegger, Martin;Palsson, Bernhard
通讯作者：
Palsson, Bernhard
影响因子：
46.9
作者：
Farmer, WR;Liao, JC
通讯作者：
Liao, JC
影响因子：
8.4
作者：
Keasling, Jay D.
通讯作者：
Keasling, Jay D.