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基于细菌的丁亚体的工程氧普林醇响应的 рибо开关用于哺乳动物细胞培养中的基因表达控制

Vera Hedwig^1,2, Maike Spöring^1,2, Julia Ottlinger¹

¹Department of Chemistry, University of Konstanz, Konstanz, Germany.

Nucleic acids research

|March 15, 2025

概括

研究人员开发了新的RNA开关,通过使用安全药物代谢物oxypurinol来控制基因表达. 这为生物工具和基因疗法提供了一种无蛋白质的方法.

科学领域:

分子生物学分子生物学
生物技术是生物技术.
在RNA治疗方面,RNA疗法.

背景情况:

带状切换器为研究和治疗提供无蛋白质的基因表达控制.
目前的合成 рибо开关具有有限的连接体选择,通常需要 de novo aptamer 选择.

研究的目的:

为了识别合成 рибо开关的新配体.
为人类细胞中基因表达控制开发oxypurinol响应的核糖开关.

主要方法:

细菌的Xanthine I riboswitch aptamer与oxypurinol的相互作用的表征.
进行X射线晶体学以确定阿普他/氧皮醇复合物的结构.
在人体细胞中构建和优化基于拼接的oxypurinol riboswitch.

主要成果:

克桑丁I рибо开关的阿巴胺结合了氧普林醇,这是阿洛普林醇的代谢物.
晶体结构揭示了oxypurinol结合,类似于xanthine.
经过优化后的 рибо开关实现了转基因表达的100倍以上的诱导.

结论:

开发了功能性氧普林醇响应的核糖开关,用于按需的基因调节.
使用已确定的药物展示了无蛋白质的基因控制策略.
生物研究和潜在的基因治疗应用的先进工具.

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Riboswitches

Riboswitches

Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
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Types of RNA

Types of RNA

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Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...

Reporter Genes

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.