与循环逆转录相结合的CRISPR-Cas12a用于放大miRNA的检测
Xi Long1, Jiacheng Li2, Tong Luo1
1College of Chemistry and Chemical Engineering, Central South University, Changsha, 410083, P. R. China. yangyanjing@csu.edu.cn.
概括
我们开发了CRISPR-CRT,这是一种量化微RNA (miRNA) 表达的敏感方法. 这种测试可检测miRNA在高特异性的子女性分子水平,使得临床分子诊断.
科学领域:
- 分子生物学分子生物学
- 生物技术是生物技术.
- 诊断 诊断 诊断 诊断
背景情况:
- 微RNA (miRNA) 表达特征分析对于理解生物过程和疾病状态至关重要.
- 精确和敏感的miRNA量化对于临床分子诊断至关重要.
研究的目的:
- 开发一种高灵敏度,特异性和多用途的方法来量化miRNA表达.
- 建立一种用于检测miRNA在子女性水平的新型检测方法.
主要方法:
- 将CRISPR-Cas12a与循环逆转录 (CRT) 结合起来,以创建CRISPR-CRT测定.
- 使用CRT将miRNA目标放大到发针RT产品中.
- 使用CRISPR-Cas12a的附带分离活动被发针RT产品激活用于检测.
主要成果:
- 实现了miRNA的女性细胞下检测极限 (LOD,0.201 fM).
- 通过两阶段序列验证过程,在检测miR-21方面表现出极好的特异性.
- 在人血清样本中成功进行了miR-21水平的敏感评估,检测了与疾病相关的样本.
结论:
- 克里斯普尔-CRT为miRNA量化提供了一种高度敏感和特定的方法.
- 该试验显示了临床分子诊断的巨大潜力,特别是使用血清样本检测疾病.
相关概念视频
CRISPR
52.5K
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...
52.5K
CRISPR and crRNAs
17.1K
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...
17.1K
CRISPR/Cas9 Genome Editing
79
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...
79


