相关实验视频
Updated: Jul 18, 2025

08:50
A Nonsequencing Approach for the Rapid Detection of RNA Editing
Published on: April 21, 2022
2.6K
检查影响MS2-ADAR1脱氨酶系统对位点导向RNA编辑的因素
Md Thoufic Anam Azad1,2, Umme Qulsum1,3, Toshifumi Tsukahara1,4
1Area of Bioscience, Biotechnology and Biomedical Engineering Research Area, Japan Advanced Institute of Science and Technology (JAIST), 1-1 Asahidai, Nomi City 923-1292, Ishikawa, Japan.
Genes
|August 26, 2023
概括
优化导向RNA长度和位置增强了对RNA 1 (ADAR1) 编辑效率起作用的腺脱氨酶. 精确的酶,导向RNA和向RNA的比率对于特定的RNA编辑至关重要.
科学领域:
- 分子生物学分子生物学
- 在RNA生物学,RNA生物学.
- 基因编辑 基因编辑
背景情况:
- 作用于RNA的腺氨酸脱氨酶 (ADARs) 是修改RNA的酶.
- ADARs具有双链RNA结合域和一个deaminase域 (DD).
- 有针对性的RNA编辑为治疗应用提供了潜力.
研究的目的:
- 研究引导RNA特征对ADAR1介导RNA编辑效率的影响.
- 在mRNA中确定导向腺转化为酶转换的最佳条件.
- 为了评估目标之外的编辑效果.
主要方法:
- 利用MS2系统和自定义指导RNAs来指导ADAR1-脱氨酶域 (ADAR1-DD) 向腺残留物.
- 转染的HEK-293细胞具有编码目标mRNA,ADAR1-DD和不同长度 (19,21,23nt) 和位置 (上游/下游) 的引导RNA的结构.
- 将停止编码子 (TAG,TAA) 量化为酸盐 (TGG) 的编辑效率,并评估了目标外的修改.
主要成果:
- 上游指导RNAs显示出比下游对应物更高的功能有效性.
- 编辑效率的最佳指导RNA长度遵循了层次结构:21 nt > 23 nt > 19 nt.
- 观察到的最高编辑效率达到16.6%;非目标编辑发生在目标站点的下游,而不是在补充区域.
- 编辑效率与去氨酶度正相关,与指导RNA度负相关.
结论:
- 特定RNA编辑效率对指导RNA长度和相对于目标部位的定位非常敏感.
- 为了实现精确的RNA编辑,需要仔细优化酶,导向RNA和点RNA之间的固态度比率.
- 了解这些参数对于开发强大的RNA编辑技术至关重要.
相关概念视频
RNA Editing
9.0K
RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
9.0K
RNA Stability
33.6K
Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
33.6K
mRNA Stability and Gene Expression
5.6K
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
5.6K
Transfer RNA Synthesis
12.0K
One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
12.0K
Chromatin Structure Regulates pre-mRNA Processing
7.0K
In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
The chromatin structure, especially...
The chromatin structure, especially...
7.0K
Nuclear Export of mRNA
7.7K
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...
7.7K

