相关实验视频
Updated: Jun 23, 2026
08:25
Identification of Circular RNAs using RNA Sequencing
Published on: November 14, 2019
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一个单一的CRISPR-RCA策略,用于超敏感和特定的circRNA检测
Xinxin Ke1, Ajuan Liang2, Chuanxia Chen3
1Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, China. hutaozd@zju.edu.cn.
概括
一种新的RETA-CRISPR方法提供了循环RNAs (circRNAs) 的快速,超敏感检测. 这种基于CRISPR/Cas12a的策略实现了高特异性和灵敏度,使得早期疾病生物标志物的识别成为可能.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 基因组学就是基因组学.
背景情况:
- 准确的循环RNA (circRNA) 检测对于临床应用至关重要.
- 挑战包括低丰度和线性RNA异构体的干扰.
- 需要创新的检测方法.
研究的目的:
- 开发一种用于circRNA检测的快速,特定和超敏感的方法.
- 应用CRISPR/Cas12a系统进行circRNA量化.
- 为circRNA生物标志物建立一个新的诊断工具.
主要方法:
- 开发了RETA-CRISPR (用CRISPR/Cas12a进行反转录-滚动圆放大).
- 利用RT-RCA进行circRNA放大,产生反向拼接接头 (BSJ) 序列.
- 采用PAM独立的Cas12a/crRNA复合体用于目标识别和附带裂解.
主要成果:
- 对于circRNAs来说,已经达到300 aM的检测极限.
- 展示了30分钟至2小时的快速检测时间.
- 在细胞系中成功检测出肝细胞癌生物标志物hsa_circ_0001445.
结论:
- 在RETA-CRISPR中,指数放大与CRISPR/Cas12a的附带活动相结合.
- 这种方法为circRNA检测提供了一个敏感和特定的工具.
- 将RETA-CRISPR定位为基于CRISPR的诊断的一个有前途的平台.
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...