接近同源的tRNAs提高了伪尿素介导的早期终结子的读透效率和精度
Nan Luo1, Qiang Huang1, Liting Dong1,2
1State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China.
Nature biotechnology
|March 6, 2024
概括
RESTART v3 增强了 RNA 基编辑功能,可以读取过早终结子 (PTC),从而改善遗传疾病模型中的蛋白质功能. 与以前的版本相比,这种方法为纠正PTC提供了更高的效率和精度.
科学领域:
- 分子生物学分子生物学
- 基因工程是一种基因工程.
- 在RNA治疗方面,RNA疗法.
背景情况:
- 过早终止密码子 (PTC) 导致非功能性蛋白质,导致遗传疾病.
- 当前的PTC抑制方法在效率和精度方面存在局限性.
研究的目的:
- 开发一个改进的RNA基编辑器,以实现高效和精确的PTC阅读.
- 在细胞和疾病模型中评估新系统的有效性.
主要方法:
- 开发RESTART v3,使用近同类tRNA用于伪尿素修饰的PTC.
- 在培养细胞和疾病模型 (囊性纤维化,赫勒综合征) 中测试RESTART v3.
- 评价编辑效率,PTC读透精度,以及非目标效应.
主要成果:
- 与RESTART v2.2相比,RESTART v3的编辑效率平均增加了约5倍.
- 在疾病细胞模型中实现了功能性PTC读透.
- 原始氨基酸的准确重新整合发生在近一半的PTC部位,而不会影响正常终止.
结论:
- RESTART v3代表了可编程RNA伪化用于PTC抑制的重大进步.
- 该系统在PTCs引起的遗传疾病中显示出治疗应用的前景.
- 虽然观察到非目标站点,但它们没有改变编码信息或转录表达水平.
相关概念视频
Improving Translational Accuracy
10.4K
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...
10.4K
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
Transfer RNA Synthesis
11.9K
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...
11.9K
Termination of Translation
25.4K
The large ribosomal subunit has several important structures essential to translation. These include the peptidyl transferase center (PTC) - which is the site where the peptide bond is formed - and a large, internal, water-filled tube through which the nascent polypeptide moves. This latter structure is called the Peptide Exit Tunnel, and it begins at the PTC and spans the body of the large ribosomal subunit. During translation, as the nascent polypeptide chain is synthesized, it passes through...
25.4K
Leaky Scanning
5.1K
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...
5.1K
tRNA Activation
19.2K
Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
19.2K


