全球对5'单酸化mRNA衰变中间体的分析和分析
Ai-Ping Chen1, Wan-Chieh Chen2, Bo-Han Hou2
1Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan.
Methods in molecular biology (Clifton, N.J.)
|November 13, 2024
概括
本研究介绍了RNA末端并行分析 (PARE),这是一种高通量测序方法,用于分析RNA衰变中间体. 在真核生物中,PARE可对mRNA降解途径进行全基因组分析.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 在RNA生物学,RNA生物学.
背景情况:
- RNA循环涉及核糖核酶产生特定5'末端的衰变中间体.
- 这些中间体是从切割,微RNA裂变或从复合体中保护的碎片中产生的.
- 研究RNA衰变途径需要对这些中间体进行全球分析.
研究的目的:
- 描述一种高通量测序协议,用于基因组范围内的RNA衰变中间体的分析.
- 为分析RNA降解组数据提供生物信息工具和脚本.
- 突出修改以优化测序质量和适应新平台的修改.
主要方法:
- 对RNA末端的并行分析 (PARE) 协议.
- 高通量测序用于全基因组分析.
- 对RNA降解组数据的生物信息分析.
主要成果:
- 启用了5'单酸化mRNA衰变中间体的全基因组分析.
- 提供用于PARE数据分析的工具和脚本.
- 详细介绍了优化的图书馆构建和生物信息分析.
结论:
- PARE是研究各种RNA衰变途径的一个有价值的方法.
- 提供的工具有助于对RNA降解组数据进行全面分析.
- 该协议可以适应不断发展的测序技术.
相关概念视频
mRNA Stability and Gene Expression
5.5K
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
5.5K
Nuclear Export of mRNA
7.6K
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...
7.6K
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
Ribosome Profiling
3.5K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.5K
Pre-mRNA Processing: Modification of pre-mRNA Ends
9.2K
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...
9.2K
Regulation of Expression at Multiple Steps
873
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...
873


