一个用于体外选择完全2'-修饰RNA体的系统
Emily D Ziperman1, Kate B Fitzpatrick1, Malavika A Nair1
1Department of Chemistry, Brandeis University, Waltham, Massachusetts, USA. kraussi@brandeis.edu.
Organic & biomolecular chemistry
|February 6, 2025
概括
这项研究表明,特定的聚合酶与SELection with Modified Aptamers (SELMA) 兼容,用于发现具有大量修饰的体. 这使得能够开发出具有增强功能的新型aptamer.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 合成生物学 合成生物学
背景情况:
- 在开发新型分子工具方面,aptamer发现方法至关重要.
- 用修改的阿普塔默 (SELMA) 进行选择,可以结合大型核基修改,如甘氨酸.
- 酶与改性核酸的兼容性是aptamer工程中的一个关键挑战.
研究的目的:
- 研究SFM4-3,KOD DGLNK和Therminator聚合酶与SELMA方法的兼容性.
- 为了使大型核基修饰,包括甘氨酸,可以使用SELMA将其纳入aptamers.
- 优化原料设计和协议,以提高SELMA性能.
主要方法:
- 使用SELection与修改的Aptamers (SELMA) 进行aptamer发现.
- 测试了SFM4-3,KOD DGLNK和Therminator聚合酶,以检测它们的纳入改性核基的能力.
- 修改了原料设计和实验协议,以促进酶兼容性.
主要成果:
- 发现SFM4-3,KOD DGLNK和Therminator聚合酶与SELMA兼容.
- 该方法成功地在所有位置上实现了2'-或2'-甲基核糖的修饰.
- 热因聚合酶证明了基因修饰核基的优越结合,用于使用2'-基修饰的点击化学.
结论:
- 优化的SELMA协议和原料设计允许使用具有改性核酸的标准DNA聚合酶.
- 这一进步扩大了通过SELMA可实现的aptamer修饰的范围.
- 热聚合酶在阿普坦体开发中显示出对点击化学应用的前景.
相关概念视频
Types of RNA
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...
RNA Interference
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...
Riboswitches
Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
Types of 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 regulating 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 Performs Diverse...
RNA Performs Diverse...
Pre-mRNA Processing: Modification of pre-mRNA Ends
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 the cell...
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 the cell...
Nucleic Acid Structure
The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group on the ribose's second carbon and a hydrogen on the deoxyribose's second carbon. The phosphate residue attaches to the hydroxyl group of the 5′ carbon of one sugar and the hydroxyl group of the 3′ carbon of the sugar of the next nucleotide, which forms a 5′ to 3′ phosphodiester linkage.
DNA Structure
DNA has a double-helix structure. The...
DNA Structure
DNA has a double-helix structure. The...


