相关实验视频
Updated: Jul 10, 2025

08:50
A Nonsequencing Approach for the Rapid Detection of RNA Editing
Published on: April 21, 2022
2.6K
使用人工MS2-APOBEC系统改进C-to-URNA编辑
Jiarui Li1, Guangyao Fan1,2, Matomo Sakari1
1Bioscience, Biotechnology and Biomedical Engineering Research Area, Japan Advanced Institute of Science and Technology, Nomi, Ishikawa, Japan.
Biotechnology journal
|November 27, 2023
概括
研究人员通过使用APOBEC3A和APOBEC3G酶设计了MS2系统,以实现RNA细胞丁脱胺 (C-to-U编辑). 这种新的方法显示了增强RNA编辑应用的潜力.
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:
- RNA细胞因子去胺,特别是C-to-U编辑,是一个关键的转录后修改.
- 在MS2-阿波利波蛋白B编辑催化多类 (APOBEC) 1系统之前已经启用了RNA编辑.
- APOBEC3A和APOBEC3G是具有已知的单链RNA偏好的细胞氨酸脱氨酶 (CDA) 酶.
研究的目的:
- 为了研究APOBEC3A和APOBEC3G与MS2系统融合的RNA编辑效率.
- 设计指导RNA,以促进APOBEC3A和APOBEC3G的单链RNA结构.
- 通过特定站点的突变改变其基准偏好来提高APOBEC3G编辑效率.
主要方法:
- APOBEC3A和APOBEC3G酶与MS2系统的融合.
- 设计非传统的引导RNA来诱导目标序列循环,创建单链RNA结构.
- 将D317W突变引入APOBEC3G以修改其基本偏好.
- 同时感染HEK 293T细胞与MS2融合酶和导向RNA.
- 对RNA编辑效率的评估.
主要成果:
- 与MS2融合的APOBEC3A和APOBEC3G在与循环诱导导向导RNA共传染时,显示出高的C-to-URNA编辑效率.
- 在APOBEC3G中D317W突变显示在编辑效率上有轻微的,但不具有统计学意义的改善.
- 在设计的引导RNA的存在下,APOBEC3A和APOBEC3G成功诱导了C-to-URNA编辑.
结论:
- APOBEC3A和APOBEC3G可以通过使用循环引导RNA的MS2系统进行C-to-URNA编辑.
- 特定站点的突变可以通过改变其基础偏好来提高APOBEC3G的编辑效率.
- MS2系统在与不同酶的融合中的多功能性表明了RNA编辑应用的广泛潜力.
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
Pre-mRNA Processing: Modification of pre-mRNA Ends
9.3K
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a cap to the 5' end of the growing transcript. In this process, a 5' phosphate is replaced by modified guanosine that has a methyl group attached (7-methyl guanosine). This 5' cap helps...
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a cap to the 5' end of the growing transcript. In this process, a 5' phosphate is replaced by modified guanosine that has a methyl group attached (7-methyl guanosine). This 5' cap helps...
9.3K
Improving Translational Accuracy
11.1K
Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
11.1K
Nonsense-mediated mRNA Decay
10.6K
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,...
10.6K
RNA Splicing
56.4K
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
56.4K
pre-mRNA Processing
52.9K
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl...
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl...
52.9K

