二次毛囊能够在难以编辑的部位进行高效的生殖线mtDNA基编辑
Qin Xie1, Haibo Wu1, Hui Long1
1Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China.
Molecular therapy. Nucleic acids
|April 1, 2024
概括
这项研究引入了一种新的生殖线线线粒体DNA (mtDNA) 编辑策略,通过注入基编辑器到二次毛囊卵细胞中. 这种方法提高了编辑效率,特别是在困难的网站,并成功地创建了活体动物模型,而不会增加目标之外的风险.
科学领域:
- 线粒体遗传学线粒体遗传学
- 基因编辑技术 基因编辑技术
- 发育生物学是发展生物学.
背景情况:
- 细菌线线线粒体DNA (mtDNA) 编辑对于疾病建模和基因治疗至关重要.
- DddA衍生型细胞因子基编辑器 (DdCBEs) 能够精确地编辑mtDNA.
- 目前的囊胚注射方法在早期开发过程中面临mtDNA复制的挑战,这限制了某些部位的编辑效率.
研究的目的:
- 开发一个改进的生殖线mtDNA基编辑策略.
- 为了克服囊胚注射的局限性,用于编辑难以编辑的mtDNA站点.
- 为了提高生殖线mtDNA编辑的效率和安全性.
主要方法:
- 向二次毛囊卵细胞注射DdCBEs,这是一个具有活跃mtDNA复制的阶段.
- 在具有挑战性的mtDNA位置使用这种策略进行G-to-A基转换.
- 从编辑的卵细胞生成活体动物模型.
主要成果:
- 在以前难以编辑的mtDNA站点实现了高效的G-to-A转换.
- 通过二次卵泡卵细胞注射方法成功生产了活体动物模型.
- 与胚胎注射相比,基础编辑效率增加了3倍.
- 证实,在二次毛囊中进行编辑,与囊注射相比,不会增加非目标效应.
结论:
- 将DdCBEs注射到二次卵泡卵子细胞是胚胎mtDNA编辑的有效策略.
- 这种方法显著提高了编辑效率,特别是对于具有挑战性的mtDNA站点.
- 该方法是安全的,没有增加非目标效应的风险,可以创建疾病模型和潜在的基因疗法.
相关概念视频
02:59Animal Mitochondrial Genetics
7.6K
Among all the organelles in an animal cell, only mitochondria have their own independent genomes. Animal mitochondrial DNA is a double-stranded, closed-circular molecule with around 20,000 base pairs. Mitochondrial DNA is unique in that one of its two strands, the heavy, or H, -strand is guanine rich, whereas the complementary strand is cytosine rich and called the light, or L, -strand. Compared to nuclear DNA, mitochondrial DNA has a very low percentage of non-coding regions and is marked by...
7.6K
02:23RNA Editing
9.0K
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...
9.0K
01:02Long-patch Base Excision Repair
7.0K
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:
7.0K
02:46Genome Copying Errors
4.2K
DNA replication is a well-evolved process that copies millions of base pairs with high fidelity during each cell division. Occasionally a wrong base or a long stretch of wrong bases may get added to the daughter strands. If the errors are left unchecked, cells might accumulate several mutations that might endanger their survival. Therefore, the copying errors are checked and repaired at three levels.
4.2K
01:20Mismatch Repair
4.8K
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...
4.8K
02:08Gene Conversion
9.7K
Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
9.7K