通过调制新合成的DNA链上的甲基化和延长表达,提高主要编辑效率
Xiaosong Han1,2, Xianghua Xu1, Youcai Xiong1
1Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education & Key Laboratory of Swine Genetics and Breeding of Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan, 430070, P. R. China.
Advanced science (Weinheim, Baden-Wurttemberg, Germany)
|March 7, 2025
概括
研究人员发现,增加DNA甲基化提高了首席编辑器 (PE) 的效率. 他们开发了一种经期元素驱动的PE系统 (epiPE2),可以提高精确基因编辑,并允许在人类细胞中进行多重编辑.
科学领域:
- 遗传学 是一个遗传学.
- 分子生物学分子生物学
- 表观遗传学 在表观遗传学中,表观遗传学是指表观遗传学.
背景情况:
- 主编辑器 (PEs) 是精确基因组工程的先进工具.
- 了解PE修复机制对于扩大其应用至关重要.
- 在此之前,DNA甲基化对PE修复途径的作用尚不清楚.
研究的目的:
- 调查CpG甲基化对主要编辑器效率的影响.
- 阐明新型DNA甲基转移酶 (DNMT3A/3B) 在PE修复中的参与.
- 开发和描述一种新的以情节元素驱动的原始编辑系统 (epiPE).
主要方法:
- 研究了CpG甲基化水平与PE效率之间的相关性.
- 确定了DNMT3A/3B在PE修复途径中的作用.
- 开发了一种使用EBNA1/oriP的经期元素驱动的PE系统 (epiPE).
- 对EPE2与已建立的PE系统 (PE2,lentiPE2,PE4max) 进行了比较分析.
- 在多个遗传位置评估了多重编辑能力.
主要成果:
- 新合成的链上增加的CpG甲基化显著提高了PE的效率.
- 新的DNA甲基转移酶 (DNMT3A/3B) 是PE修复途径的组成部分.
- 与PE2相比,新型epiPE2系统的编辑效率提高了2.0至38.2倍.
- epiPE2保持了最小的插入和删除 (indel) 速率.
- 在人类细胞中,epiPE2系统实现了高效的多重精确基因编辑,最高可在10个位置进行编辑.
结论:
- 这项研究加深了对主要编辑修复机制的理解.
- 开发的epiPE2系统为主要编辑提供了更高的效率和复杂化功能.
- epiPE2为基因组工程的基础研究和临床应用提供了一个有前途的工具.
相关概念视频
Chromatin Modification in iPS Cells
1.6K
Chromatin modification alters gene expression; therefore, scientists can add histone-modifying enzymes, histone variants, and chromatin remodeling complexes to somatic cells to aid reprogramming into pluripotent stem (iPS) cells.
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
1.6K
Epigenetic Regulation
3.0K
Epigenetic changes alter the physical structure of the DNA without changing the genetic sequence and often regulate whether genes are turned on or off. This regulation ensures that each cell produces only proteins necessary for its function. For example, proteins that promote bone growth are not produced in muscle cells. Epigenetic mechanisms play an essential role in healthy development. Conversely, precisely regulated epigenetic mechanisms are disrupted in diseases like cancer.
X-chromosome...
X-chromosome...
3.0K
Chromatin Structure Regulates pre-mRNA Processing
6.9K
In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
The chromatin structure, especially...
The chromatin structure, especially...
6.9K
Spreading of Chromatin Modifications
8.2K
The histone proteins in the nucleosomes are post-translationally modified (PTM) to increase or decrease access to DNA. The commonly observed PTMs are methylation, acetylation, phosphorylation, and ubiquitination of lysine amino acids in the histone H3 tail region. These histone modifications have specific meaning for the cell. Hence, they are called "histone code". The protein complex involved in histone modification is termed as "reader-writer" complex.
Writers
The writer...
Writers
The writer...
8.2K
Proofreading
6.1K
Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore, it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase...
Errors During Replication are Corrected by the DNA Polymerase...
6.1K
Regulation of Expression at Multiple Steps
862
The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
862


