通过使用工程可光切换RNA结合蛋白来控制RNA代谢和CRISPR-Cas功能的时空控制
Renmei Liu1,2,3, Jing Yao1,2, Siyu Zhou1,2
1Optogenetics & Synthetic Biology Interdisciplinary Research Center, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
Nature protocols
|November 30, 2023
概括
研究人员开发了LicV,一种光激活的合成RNA结合蛋白 (RBP),可以精确控制RNA功能. 这项技术提供了毫秒的时空分辨率,用于研究RNA代谢和基因编辑.
科学领域:
- 分子生物学分子生物学
- 视觉遗传学 视觉遗传学
- 在RNA生物学,RNA生物学.
背景情况:
- RNA分子是细胞过程的核心,包括基因表达和催化.
- RNA结合蛋白 (RBPs) 调节RNA功能,但时间空间控制方法是有限的.
- 光遗传工具通过使用光来精确控制生物过程.
研究的目的:
- 开发一种合成的可光开关RNA结合蛋白 (RBP) 用于RNA的光遗传控制.
- 为设计可光切换RBPs和适应CRISPR-Cas系统建立一个可通用的协议.
- 为了实现精确的RNA代谢和基因编辑的时空调节.
主要方法:
- 开发LicV,一种由蓝光激活的合成可光切换RBP.
- 应用LicV用于RNA局部化,拼接,翻译和稳定性的光遗传控制.
- 通过基于LicV的策略,调整CRISPR-Cas系统用于光遗传控制.
主要成果:
- LicV成功实现了各种RNA相关细胞过程的光遗传调节.
- 基于LicV的系统展示了高动态范围和亚微米/毫秒的时空分辨率.
- 为了设计可光切换的RBP和转换CRISPR-Cas系统,建立了一个协议.
结论:
- LicV为RNA代谢的光遗传控制提供了一种高效和可通用的策略.
- 开发的协议有助于为RNA和CRISPR-Cas功能创建新的可光切换效应器.
- 这种方法在扰乱细胞过程的传统方法上具有优势.
相关概念视频
01:23Types of RNA
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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.
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01:56Riboswitches
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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...
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01:23RNA Interference
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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...
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02:15Experimental RNAi
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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...
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02:41Chromatin Structure Regulates pre-mRNA Processing
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The chromatin structure, especially...
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01:59CRISPR
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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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