在Saccharomyces cerevisiae中,mRNA和小分子触发条件导向RNA失活
Chenggang Xi1, Stephen Chiu2, William E Voje3
1Department of Energy, Environmental and Chemical Engineering, Washington University in St. Louis, St. Louis, MO 63130, United States.
New biotechnology
|July 18, 2025
概括
这项研究引入了可切换导向RNA (gRNA) 用于动态CRISPR干扰 (CRISPRi) 基因控制. 这些工程化gRNAs使可调节的基因表达能够响应mRNA或小分子信号,从而推进生物工程应用.
科学领域:
- 合成生物学 合成生物学
- 分子生物学分子生物学
- 生物工程是生物工程.
背景情况:
- 克里斯普尔干扰 (CRISPRi) 提供精确的基因表达调制,但缺乏动态控制.
- 现有的CRISPRi系统在很大程度上是静态的,限制了实时基因调节.
- 动态控制对于复杂的生物应用,如代谢工程,至关重要.
研究的目的:
- 开发可切换导向RNA (gRNA) 用于使用CRISPRi进行动态和可调节的基因表达控制.
- 为了使CRISPRi调节能够响应外部信号,如mRNA或小分子.
- 为基于gRNA的生物计算和可逆基因控制创建一个模块化平台.
主要方法:
- 设计了具有5'或3'扩展的gRNAs来阻止CRISPRi的功能.
- 使用托管介导的链位移和aptzymes用于响应信号的gRNA失活.
- 使用mRNA和小分子触发器证明了gRNA沉默和恢复功能.
- 在Saccharomyces cerevisiae中实现模块化gRNA用于遗传逻辑门的构建.
主要成果:
- 通过设计的扩展实现了gRNA活动的完全沉默.
- 通过信号诱导的分离或裂变成功恢复了gRNA功能.
- 通过特定的mRNA和小分子信号来证明工程化gRNA的非激活.
- 在酵母中建立了基于gRNA的生物计算与多输入遗传逻辑门.
结论:
- 开发了一种使用工程化gRNAs进行动态和可逆的基因表达控制的新策略.
- 可切换的gRNA系统提供可调节的CRISPRi调节,以响应细胞或环境信号.
- 这种方法扩大了高级合成生物学和真核生物系统中的生物计算工具包.
相关概念视频
02:51mRNA Stability and Gene Expression
5.7K
The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
Cis-acting Elements involved in mRNA stability
5.7K
02:27Nonsense-mediated mRNA Decay
10.8K
The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
10.8K
01:23RNA Interference
26.5K
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...
26.5K
01:29Translational Regulation
99
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,...
99
01:23Types of RNA
65.0K
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...
65.0K
02:31Nuclear Export of mRNA
7.9K
Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
7.9K