来自宿主转录的非正规crRNAs可以通过Cas9检测多重化RNA
Chunlei Jiao1, Sahil Sharma2, Gaurav Dugar2
1Helmholtz Institute for RNA-based Infection Research (HIRI)/Helmholtz-Centre for Infection Research (HZI), 97080 Würzburg, Germany.
Science (New York, N.Y.)
|April 28, 2021
概括
科学家设计了一个CRISPR-Cas系统,通过创建非正规的CRISPRRNA (crRNA) 来检测RNA. 这导致了LEOPARD诊断平台用于多重RNA检测,包括SARS-CoV-2变种.
科学领域:
- 分子生物学
- 克里斯普尔-卡斯系统
- 诊断技术
背景情况:
- CRISPR-Cas系统使用CRISPRRNAs (crRNAs) 来识别外来遗传物质.
- 类型II CRISPR-Cas系统依赖转激活crRNA (tracrRNA) 来处理crRNA和利用Cas9酶.
- 通常与特定的crRNA杂交.
研究的目的:
- 研究来自Campylobacter jejuni的Cas9-RNA复合物的相互作用.
- 发现新的crRNA形成机制及其针对DNA的潜力.
- 根据修改的tracrRNA设计一个多功能RNA检测平台.
主要方法:
- 来自Campylobacter jejuni的Cas9-RNA复合物的分析.
- 发现了向内源细胞RNA杂交的tracrRNA,形成非正规的crRNA.
- 重编程的tracrRNAs的工程,以将RNA存在与Cas9介导的DNA向联系起来.
- 开发LEOPARD平台 (利用工程化tracrRNA和目标DNA进行并行RNA检测).
主要成果:
- 通过向细胞RNA混合生成的非正规crRNAs,能够指导Cas9DNA向.
- 根据任何所需的RNA的存在,设计的tracrRNAs可实现DNA向.
- 在单个测试中,LEOPARD平台同时检测了多个病毒RNA.
- 在患者样本中,LEOPARD以单核酸精度区分了SARS-CoV-2及其D614G变体.
结论:
- 非正规crRNA形成的发现扩大了对CRISPR-Cas系统的理解.
- 设计的tracrRNA为RNA指导的DNA定位提供了一种新的机制.
- 利奥帕德平台为敏感和特定RNA检测,包括病毒诊断提供了一种强大,可复合的工具.
相关概念视频
01:59CRISPR
54.2K
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...
54.2K
02:53CRISPR and crRNAs
18.0K
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...
18.0K
03:21RNA-seq
10.9K
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
10.9K
01:28CRISPR/Cas9 Genome Editing
829
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...
829
02:31Homologous Recombination
58.3K
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...
58.3K
01:23RNA Interference
27.0K
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
27.0K