Scholarlink

<p>Cas13a-based diagnostic systems have been widely utilized for the detection of RNA targets. However, without preamplification such systems are difficult to realize ultrasensitive (RT-PCR level) single pot RNA detection. Here, we found that double strand RNA can effectively activate the <i>trans</i>-cleavage activity of Cas13a RNP, while the cleavage rates of dsRNA by activated Cas13a RNP are very low. In addition, specially designed RNA-Nanocircle has limited ability to activate Cas13a RNP, but this activation is restored once the circular structures are cleaved and become linear. Based on this original method to control <i>trans</i>-cleavage activity of Cas13a RNP, we developed a Cas13a autocatalytic biosensing system assisted by RNA-Nanocircles, which allows one target RNA to activate numerous Cas13a RNPs. With this approach we show ultrasensitive detection of 1aM of synthetic RNA targets without preamplification within 15 min. The clinical utility of this biosensor was validated by monitoring miRNA-21 levels in plasma samples from colorectal cancer patients. This innovative approach highlights the potential of Cas13a-based biosensors in precision oncology, offering a rapid, non-invasive, and ultrasensitive method for RNA biomarker detection in liquid biopsies.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Autocatalytic Cas13a biosensor enabled by RNA-nanocircles for ultrasensitive RNA detection

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年1月13日

查看全文
  • Fei Deng,
  • Sneha Gulati,
  • Rui Sang,
  • Yi Li,
  • Ewa M. Goldys

摘要

Cas13a-based diagnostic systems have been widely utilized for the detection of RNA targets. However, without preamplification such systems are difficult to realize ultrasensitive (RT-PCR level) single pot RNA detection. Here, we found that double strand RNA can effectively activate the trans-cleavage activity of Cas13a RNP, while the cleavage rates of dsRNA by activated Cas13a RNP are very low. In addition, specially designed RNA-Nanocircle has limited ability to activate Cas13a RNP, but this activation is restored once the circular structures are cleaved and become linear. Based on this original method to control trans-cleavage activity of Cas13a RNP, we developed a Cas13a autocatalytic biosensing system assisted by RNA-Nanocircles, which allows one target RNA to activate numerous Cas13a RNPs. With this approach we show ultrasensitive detection of 1aM of synthetic RNA targets without preamplification within 15 min. The clinical utility of this biosensor was validated by monitoring miRNA-21 levels in plasma samples from colorectal cancer patients. This innovative approach highlights the potential of Cas13a-based biosensors in precision oncology, offering a rapid, non-invasive, and ultrasensitive method for RNA biomarker detection in liquid biopsies.