由基编辑器引起的非目标RNA编辑热点
Michelle Shmuel-Eidelman1, Roni Cohen-Fultheim1, Eli Eisenberg2
1Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat Gan 5290002, Israel; The Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat Gan 5290002, Israel.
Molecular therapy : the journal of the American Society of Gene Therapy
|December 25, 2025
概括
这项研究引入了一种新方法来检测基因编辑器,基因编辑工具的意外RNA编辑. 它可以识别特定的基因和易受这些非目标编辑的区域,这对于开发更安全的基因疗法至关重要.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 生物技术是生物技术.
背景情况:
- 基准编辑器为治疗应用提供精确的基因组和转录组修改.
- 基准编辑的临床翻译受到非目标,无意编辑的限制.
- 目前的方法主要检测反复发生的RNA脱胺,错过的随机脱事件.
研究的目的:
- 开发和应用一种新的方法来量化基因编辑器诱导的RNA变化的总负担.
- 识别特定的基因和基因组区域易受非目标基因编辑.
- 增强对基因编辑器特异性的理解,并指导开发更安全的基因编辑工具.
主要方法:
- 应用RNA编辑索引算法来量化非目标RNA的变化.
- 系统分析跨个体基因和外子区域的非目标编辑水平.
- 识别热点基因和局部区域容易发生异常基因编辑.
主要成果:
- 使用RNA编辑指数对基因间非目标RNA编辑水平的量化.
- 鉴定了2,844个腺因基编辑器和1,253个细胞因基编辑器热点基因.
- 发现局部异构子区域易受非目标编辑,包括那些引入过早停止编码子的区域.
结论:
- 开发的方法有效地捕获了常规检测错过的随机非目标编辑.
- 了解非目标景观对于评估治疗风险至关重要,例如过早停止子引入.
- 这项研究为提高基因编辑器精度和加速开发更安全的基因编辑技术提供了一个框架.
相关概念视频
RNA Editing
9.7K
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...
9.7K
Nonsense-mediated mRNA Decay
11.6K
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,...
11.6K
Mismatch Repair
6.2K
Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
6.2K
Mismatch Repair
43.4K
Overview
43.4K
Long-patch Base Excision Repair
7.8K
Since the discovery of the two BER pathways, there has been a debate about how a cell chooses one pathway over the other and the factors determining this selection. Numerous in vitro experiments have pointed out multiple determinants for the sub-pathway selection. These are:
7.8K
Improving Translational Accuracy
14.0K
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...
14.0K


