Harnessing crRNA Transformer for Facile and Specific Nucleic Acid Detection

CRISPR/Cas systems have emerged as promising tools for nucleic acid detection. However, their practical applications have been limited by poor specificity and the need for additional preprocessing steps. Inspired by the concept of transformers, we found that changing the forms of crRNA with spatial arrangement may endow CRISPR/Cas with an enhanced performance for nucleic acid detection. Specifically, we rationally designed two crRNA transformers─swap crRNA and split crRNA─and found that they direct the CRISPR/Cas system for <i>cis</i>- and <i>trans</i>- cleavage with enhanced specificity and decreased Cas binding affinity and possess both DNA and RNA detection abilities. Based on these findings, our strategy enabled the identification of clinical prostatic cancer in a one-step reaction, with a remarkable sensitivity of 90.0% and specificity of 96.0%. Our study deepens the understanding of CRISPR/Cas systems and introduces a promising strategy for simple nucleic acid detection with enhanced specificity, sensitivity, and functionality in clinical molecular diagnosis.

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Publication Details

Journal
Analytical Chemistry
Published
2025-02-05
DOI
https://doi.org/10.1021/acs.analchem.4c05399
Citations
14
Primary Topic
CRISPR and Genetic Engineering
Type
article
Field-Weighted Citation Impact
11.28

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article

Harnessing crRNA Transformer for Facile and Specific Nucleic Acid Detection

Xueyuan Huang, Mei Chen, Jinrong Min, Zhaoxin Huang et al.
14 citations
Analytical Chemistry
CRISPR and Genetic Engineering
11.28
article

Harnessing crRNA Transformer for Facile and Specific Nucleic Acid Detection

Xueyuan Huang, Mei Chen, Jinrong Min, Zhaoxin Huang, Guoliang Ke, Jialin Wen, Chaoyong James Yang, Mei Wen, Peng Zhang, Xiaobing Zhang, Ting Chen, Yue Ding, Yin Wang, Menghui Wang, Bo Wang
article en
14 citations

Abstract

CRISPR/Cas systems have emerged as promising tools for nucleic acid detection. However, their practical applications have been limited by poor specificity and the need for additional preprocessing steps. Inspired by the concept of transformers, we found that changing the forms of crRNA with spatial arrangement may endow CRISPR/Cas with an enhanced performance for nucleic acid detection. Specifically, we rationally designed two crRNA transformers─swap crRNA and split crRNA─and found that they direct the CRISPR/Cas system for <i>cis</i>- and <i>trans</i>- cleavage with enhanced specificity and decreased Cas binding affinity and possess both DNA and RNA detection abilities. Based on these findings, our strategy enabled the identification of clinical prostatic cancer in a one-step reaction, with a remarkable sensitivity of 90.0% and specificity of 96.0%. Our study deepens the understanding of CRISPR/Cas systems and introduces a promising strategy for simple nucleic acid detection with enhanced specificity, sensitivity, and functionality in clinical molecular diagnosis.

Analytical ChemistryVol. 97(7)
Central South University (CN), Hunan University (CN), Shanghai Jiao Tong University (CN), Xiamen University (CN), Renji Hospital (CN), Division of Materials Science and Engineering (AU), Institute of Molecular Medicine (IN), Third Xiangya Hospital (CN)
National Natural Science Foundation of China, China Postdoctoral Science Foundation, Natural Science Foundation of Hunan Province
Openalex Percentile: Top 1%
CRISPR and Genetic Engineering
11.28
DOISourceOpenAlex
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