使用CRISPR-Cas13a/C2c2进行核酸检测
Jonathan S Gootenberg1,2,3,4,5, Omar O Abudayyeh1,2,3,4,6, Jeong Wook Lee7
1Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
Science (New York, N.Y.)
|April 15, 2017
概括
一种新的基于CRISPR的诊断 (CRISPR-Dx) 称为SHERLOCK, 提供快速,高度敏感的核酸检测. 这种分子工具可以识别特定的病毒,细菌和突变,
科学领域:
- 分子生物学
- 生物技术
- 遗传学
背景情况:
- 核酸检测对于诊断,病原体识别和疾病监测至关重要.
- CRISPR-Cas13a系统提供了具有辅助酶活性的RNA向能力.
- 现有的方法可能缺乏对点护理应用所需的速度,灵敏度或特异性.
研究的目的:
- 开发一个快速,廉价,高度灵敏的核酸检测平台.
- 为诊断目的利用Cas13a的附带活动.
- 创建一种用于病原体检测,基因型和突变识别的多功能工具.
主要方法:
- 结合Cas13a的附带核糖酶活动与同热放大.
- 开发了一种基于CRISPR的诊断平台,名为SHERLOCK (特定高灵敏性酶记者解锁).
- 使用SHERLOCK检测特定的病毒株 (寨卡,登革热),细菌,人类DNA和无细胞瘤DNA突变.
主要成果:
- 在核酸检测中实现了原子分子灵敏度和单基不匹配特异性.
- 证明了夏洛克能够检测各种目标, 包括病毒,细菌和人类DNA.
- 展示了冷化试剂的潜力,以实现冷链独立性和纸张上的现场部署.
结论:
- 谢洛克提供了基于CRISPR的强大诊断工具,用于快速和敏感的分子检测.
- 该平台的多功能扩展到病原体识别,遗传分析和癌症突变检测.
- 软化和基于纸张的复制使SHERLOCK可用于可访问,照顾点和现场应用.
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...