构造特异性RNA体用于正常威兰德因子和2B型威兰德病之间的表型区别
Venkata R Machha1,2, Alexander Tischer1,2, Laurie Moon-Tasson1
1Division of Hematology, Department of Internal Medicine, Mayo Clinic, 200 1st St SW, Rochester, MN, USA.
NAR molecular medicine
|December 25, 2024
概括
研究人员开发了RNA吸附体,以区分正常的·威尔布兰德因子 (VWF) 和2B型VWF变体. 这些向体向错误折叠的A1域,显示出诊断功能获取VWD表型的潜力.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 血液学 血液学 血液学
背景情况:
- ·威尔布兰德因子 (VWF) A1域通过与血小板GPIbα受体结合来启动凝血.
- 在VWF A1域中的突变可以通过损害 (2M) 或增强 (2B) 血小板粘附引起威兰德病 (VWD).
- 2B型VWD涉及功能的增益突变,这些突变破坏了A1域的稳定性和错误折叠,产生了独特的构造.
研究的目的:
- 开发特定的分子工具来区分正常的VWF与2B型VWD中的病态VWF形状.
- 为了利用2B型VWD中错误折叠的A1域作为aptamer选择的目标.
- 评估能够区分正常和VWD特异性VWF形状的aptamer的诊断潜力.
主要方法:
- 选择核酶耐药的2'-烯胺基RNA胺基对2B型V1314D型VWF A1变异在糖化A1A2A3片段中的选择.
- 使用重组VWF碎片,血VWF,VWF缩物和患者血,表征阿巴胺结合和抑制A1-GPIbα相互作用.
- 利用表面等离子体共振和基于剪切应力的血小板粘附测试来评估阿普坦特异性和功能.
主要成果:
- 分离了两个RNA亚体,W9和V1,具有选择性结合和抑制能力.
- W9专门识别了正常的VWF (WT A1A2A3),而V1针对的是2B型V1314D A1变种.
- 亚体对复合标,血VWF,VWF缩物和患者血具有R1306W型2BVWD变体的特异性.
结论:
- 正常VWF和2B型VWD蛋白的独特构造是由W9和V1体的特异性证实.
- 这些体可以区分正常的血衍生VWF和2B型VWD中发现的VWF.
- 开发的体对临床诊断严重的功能获取VWD表型具有前途.
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 Splicing
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...
Bacterial RNA Polymerase
Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Transfer RNA Synthesis
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...
RNA Editing
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...
Leaky Scanning
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA. Marilyn Kozak discovered that the sequence RCCAUGG (where R stands for...


