相关实验视频
Updated: Jan 15, 2026

11:53
Substrate Generation for Endonucleases of CRISPR/Cas Systems
Published on: September 8, 2012
28.0K
克里斯普尔-Cas12a REC2-Nuc 相互作用驱动目标链切割并限制跨切割
Anthony Newman1, Aakash Saha2, Lora Starrs1
1The Shine-Dalgarno Centre for RNA Innovation, Division of Genome Sciences and Cancer, The John Curtin School of Medical Research, The Australian National University, Canberra, ACT 2601, Australia.
Nucleic acids research
|October 9, 2025
概括
克里斯普尔-Cas12a酶表现出不同的DNA切割能力. 了解这些 cis 和 trans 分裂的差异是优化基因组编辑和 DNA 检测技术的关键.
科学领域:
- 分子生物学分子生物学
- 生物技术是生物技术.
- 遗传学 遗传学是一种遗传学.
背景情况:
- 克里斯普尔-Cas12a酶在cis和trans中进行RNA引导的DNA裂变.
- 虽然cis分离用于基因组编辑,但trans分离可以检测DNA.
- Cas12a ортолог表现出保存的结构,但具有不同的功能功效.
研究的目的:
- 调查Cas12a ортолог之间驱动明显的cis和trans切割活动的因素.
- 为了将这些裂变特性与它们在基因组编辑和DNA检测中的应用联系起来.
主要方法:
- 对三种Cas12a正规体 (FnCas12a,LbCas12a,AsCas12a) 的比较分析.
- 整合了体外DNA裂变动力学,分子动力学模拟和体内实验 (大肠杆菌等离子体干扰,人类细胞基因组编辑) 的整合.
- 产生和测试REC2和Nuc突变,包括Nuc-loop.
主要成果:
- 在Cas12a ортолог中观察到cis裂变动学的显著变化.
- 动态的REC2-Nuc相互作用似乎影响了 cis 分裂效率.
- 核环的完整性对于Cas12a的功能至关重要.
结论:
- 在Cas12a ortologue中, cis/trans 分裂的差异与特定的蛋白质相互作用和结构元素有关.
- 这项研究为优化Cas12a用于基因组编辑和DNA检测提供了洞察力.
- 这些发现突出了推进基于Cas12a的生物技术的关键特性.
相关概念视频
CRISPR/Cas9 Genome Editing
1.7K
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
1.7K
CRISPR
57.5K
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
57.5K
CRISPR and crRNAs
18.7K
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
18.7K
Conservative Site-specific Recombination and Phase Variation
6.6K
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...
6.6K
Homologous Recombination
62.6K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
62.6K
Restarting Stalled Replication Forks
6.3K
DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...
6.3K

