TGIRT-seq对tRNA衍生RNA和相关RNA修饰进行分析
Abigail Grace Johnston1, Monima Anam1, Anindya Dutta2
1Department of Genetics, University of Alabama at Birmingham, Birmingham, AL, United States.
Methods in enzymology
|February 14, 2025
概括
tRNA衍生的RNAs,调节细胞功能的小分子,在体液中丰富. 使用TGIRT测序检测的修饰tRNA衍生RNA,提供了对生物过程和潜在生物标志物的洞察力.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 生物化学 生化学
背景情况:
- tRNA衍生RNA是从tRNA中处理的小调节RNA.
- 它们在基因沉默,蛋白质合成,应激反应和遗传中发挥作用.
- 这些RNAs在体液中丰富,表明它们有可能作为生物标志物.
研究的目的:
- 描述一种用于分析tRNA衍生RNA及其修改的协议.
- 为了应对因逆转录酶停滞而导致的检测改性RNA的挑战.
主要方法:
- 使用热稳定II组内逆转录酶 (TGIRT) 来克服由RNA修饰引起的逆转录酶停滞.
- 从纯化的细胞或组织RNA生成小RNA测序库.
- 使用TGIRT引入不匹配签名,用于修改站点的基本分辨率映射.
主要成果:
- TGIRT可以检测和分析修饰的tRNA衍生RNAs.
- 该协议允许精确地绘制RNA修饰站点的地图.
- 这种方法绕过了标准小RNA测序中的偏差.
结论:
- 基于TGIRT的测序是研究tRNA衍生RNA及其修改的一个有价值的工具.
- 这种方法增强了对RNA修饰的调节作用的理解.
- 该协议有助于从体液中发现新的生物标志物.
相关概念视频
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
RNA-seq
9.8K
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
9.8K
Transfer RNA Synthesis
11.8K
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...
11.8K


