嵌入域的Nme2Cas9腺因基编辑器具有改进的活动和准范围
Nathan Bamidele1, Han Zhang1, Xiaolong Dong2
1RNA Therapeutics Institute, University of Massachusetts Chan Medical School, Worcester, MA, 01605, USA.
Nature communications
|February 17, 2024
概括
研究人员设计了Nme2Cas9 (一种基因组编辑工具),以增强其活动和准范围,用于基因编辑. 这些改进的Nme2Cas9基编辑器对于治疗应用是有效的,可以通过单个腺相关病毒载体传递.
科学领域:
- 分子生物学分子生物学
- 基因编辑技术的技术
背景情况:
- Nme2Cas9是一个紧的基因组编辑平台,以精度和广泛准而闻名.
- 现有的Nme2Cas9腺基编辑器 (Nme2-ABE) 在活动和编辑范围上有局限性.
研究的目的:
- 设计Nme2Cas9以增加紧的Nme2Cas9基础编辑器的活动和目标范围.
- 开发具有增强治疗潜力的单个AAV可交付的腺基编辑器.
主要方法:
- 域插入工程用于重新定位Nme2Cas9.9中的除氨酶域.
- 将Nme2Cas9 PAM交互域与SmuCas9的交换以扩大准范围.
- 在各种细胞类型中测试工程Nme2Cas9变体,并用于体内输送.
主要成果:
- 与N-终端融合的Nme2-ABE相比,域嵌入的Nme2Cas9变体显示了转移的编辑窗口和增加的活动.
- 通过结合SmuCas9 PAM交互域实现了扩展编辑范围.
- 在不同类型的细胞中成功引入了治疗相关的编辑.
- 工程Nme2-ABEs在体内被验证为单个AAV的交付.
结论:
- 工程Nme2Cas9基础编辑器提供增强的活动和更广泛的准能力.
- 这些进展有助于开发更有效的基因编辑工具,用于治疗应用.
- 增强的Nme2-ABEs的单个AAV输送对体内基因组编辑策略具有前景.
相关概念视频
RNA 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
Mismatch 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
Proofreading
54.1K
Overview
54.1K
Long-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
Conservative Site-specific Recombination and Phase Variation
6.0K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
6.0K
DNA Base Pairing
27.4K
Erwin Chargaff’s rules on DNA equivalence paved the way for the discovery of base pairing in DNA. Chargaff’s rules state that in a double-stranded DNA molecule,
27.4K


