用于调节哺乳动物细胞和动物中的RNA功能的小分子Aptamer
Keisuke Fukunaga1, V Dhamodharan1, Nao Miyahira1
1Nucleic Acid Chemistry and Engineering Unit, Okinawa Institute of Science and Technology Graduate University, Onna, Okinawa 904 0495, Japan.
Journal of the American Chemical Society
|March 30, 2023
概括
研究人员使用新型RNA体开发了用于哺乳动物细胞基因调节的合成体开关. 这些核突开关在低度下对ASP7967等小分子作出反应,使细胞培养和体内基因表达得到精确控制.
科学领域:
- 合成生物学
- 分子生物学
- 生物技术
背景情况:
- 合成的核突开关为生物技术和医学提供了潜力,但在哺乳动物细胞中,它们的功能有限.
- 现有的系统需要高的小分子度,阻碍实际应用.
研究的目的:
- 开发用于哺乳动物细胞中小分子介导基因调节的新型RNA胺和核糖开关.
- 在较低的小分子度下实现基因调节.
- 展示开发的核突切换器的体内应用.
主要方法:
- 在实验室中对小分子ASP7967进行RNA的选择.
- 在HEK293细胞中设计和测试基于aptazyme的 рибо开关.
- 在小鼠体内使用腺相关病毒 (AAV8) 载体来调节人类红蛋白的表达.
- 酸酶和异子跳转的酸开关机制的组合,以加强调节.
主要成果:
- 选择了一种功能性RNA胺基 (AC17-4) 并使用它来制造基于胺基的胺基开关.
- 在低至5μM的度下,Riboswitches激活了ASP2905或ASP7967的基因表达 (> 10倍).
- 在小鼠中通过口服ASP7967实现了人体红蛋白表达的体内调节.
- 一个结合的aptzyme和异子跳转的riboswitch系统实现了~300的ON/OFF比率与低基底表达.
结论:
- 在哺乳动物细胞中开发的新型合成核突变器使用了aptamer选择.
- 这种新系统可以在显著较低的小分子度下调节基因.
- 在体内证明的有效性和高性能基因控制的潜力突出显示了该系统的生物医学和生物技术潜力.
相关概念视频
Types of RNA
64.2K
Overview
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...
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...
64.2K
Experimental RNAi
6.2K
RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
6.2K
Riboswitches
8.2K
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.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
8.2K
RNA Interference
26.2K
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
26.2K
Regulation of Expression at Multiple Steps
961
The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
961
Translational Regulation
51
Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
51


