Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge CB2 3EA, U.K.
Glanbia Performance Nutrition Canada Inc., 3500 Lacey Road, Suite 1200, Downers Grove, Illinois 60515, United States.
ACS Synth Biol. 2020 Jun 19;9(6):1406-1417. doi: 10.1021/acssynbio.0c00082. Epub 2020 Jun 4.
Riboswitches are RNA regulatory elements that bind specific ligands to control gene expression. Because of their modular composition, where a ligand-sensing aptamer domain is combined with an expression platform, riboswitches offer unique tools for synthetic biology applications. Here we took a mutational approach to determine functionally important nucleotide residues in the thiamine pyrophosphate (TPP) riboswitch in the THI4 gene of the model alga , allowing us to carry out aptamer swap using THIC aptamers from and . These chimeric riboswitches displayed a distinct specificity and dynamic range of responses to different ligands. Our studies demonstrate ease of assembly as 5'UTR DNA parts, predictability of output, and utility for controlled production of a high-value compound in . The simplicity of riboswitch incorporation in current design platforms will facilitate the generation of genetic circuits to advance synthetic biology and metabolic engineering of microalgae.
Riboswitches 是一类 RNA 调控元件，通过结合特定配体来控制基因表达。由于其模块化组成，即配体感应适体结构域与表达平台相结合，riboswitches 为合成生物学应用提供了独特的工具。在这里，我们采用突变方法确定了模型藻类 THI4 基因中硫胺素焦磷酸（TPP）riboswitch 中的功能重要核苷酸残基，从而能够使用来自 和 的 THIC 适体进行适体交换。这些嵌合 riboswitches 对不同配体表现出明显的特异性和响应动态范围。我们的研究表明，作为 5'UTR DNA 部分易于组装，输出可预测，并且可用于控制 在 中的高价值化合物的生产。riboswitch 在现有设计平台中的简单集成将有助于产生遗传回路，以推进合成生物学和微藻的代谢工程。
