一个基于CRISPR-Cas12a的新型系统,具有强大的兼容性和稳定性,用于检测甘黄叶病毒
Ting Wang1, Anzhen Li1, Hong Zhao1
1Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture and Rural Affairs, National Engineering Research Center for Sugarcane, College of Agriculture, Fujian Agriculture and Forestry University, Fuzhou, China.
Microbiology spectrum
|August 9, 2024
概括
一种新的RT-MIRA-CRISPR-Cas12a方法使用原始叶子提取物快速检测到甘黄叶病毒 (SCYLV). 这种灵敏而稳定的系统为甘黄叶病提供了快速的现场检测解决方案.
科学领域:
- 植物病理学 植物病理学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 甘黄叶病 (SCYLD),由甘黄叶病毒 (SCYLV) 引起,显著降低作物产量.
- 现有的SCYLV检测方法可能耗时,需要复杂的样本准备.
- 对于SCYLD,急需快速,灵敏和可在现场部署的诊断工具.
研究的目的:
- 开发和验证一种新的SCYLV.快速检测系统.
- 将反转录-多酶同热速放大 (RT-MIRA) 与CRISPR-Cas12a技术相结合.
- 为了建立一个用户友好和敏感的方法在现场SCYLV诊断.
主要方法:
- 开发了一个单管RT-MIRA-CRISPR-Cas12a测定.
- 使用原始甘叶提取物直接作为模板,消除了RNA提取的需要.
- 使用现场采集的样本与传统RT-PCR和RT-qPCR验证了试验的性能.
主要成果:
- 该RT-MIRA-CRISPR-Cas12a系统表现出高特异性和灵敏度,与RT-qPCR相美.
- 达到了SCYLV.LV的25个副本的低检测极限.
- 完成了整个检测过程,从样品到结果,仅需52-57分钟.
- 结果是肉眼可视检测到的.
- 该试验显示出良好的稳定性,在4°C下一周后可以检测到阳性样本.
结论:
- RT-MIRA-CRISPR-Cas12a方法为SCYLV检测提供了一个快速,灵敏和稳定的方法.
- 它使用原始叶子提取物的能力简化了样本处理,使其成为现场诊断的理想选择.
- 这一创新系统解决了对有效工具的迫切需要,以管理甘黄叶病.
相关概念视频
01:59CRISPR
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 Short...
02:53CRISPR and crRNAs
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...
01:59CRISPR
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 Short...
01:23The Antiviral System of Bacteria and Archaea: CRISPR
CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this defense.
01:28CRISPR/Cas9 Genome Editing
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...