PRICE: direct and robust detection of microRNAs at single-nucleotide resolution
Accurate single-nucleotide discrimination of miRNA is clinically vital because small sequence variations can have significant phenotypic and clinical consequences, yet existing techniques can only detect single nucleotide variations (SNVs) at specific loci. Here, we present a generalized peptide nucleic acid (PNA) mediated CRISPR/Cas13a system (PRICE), enabling detection of SNVs in miRNA sequence without sacrificing the sensitivity. PRICE utilizes PNA blockers fully complementary to non-target miRNAs (e.g., miRNAs containing SNVs at loci of no interest) but not to the target miRNA. These blockers selectively hybridize with and inhibit non-target sequences in samples (serum, cells, or tissues). Only the unhybridized target miRNA then binds to crRNA within the Cas13a complex, activating Cas13a to cleave a fluorescent reporter-quencher linker, generating a detectable signal (~10 fM limit). By designing a panel of PNAs against SNVs, PRICE provides a versatile, amplification-free platform for precise miRNA analysis, advancing cancer diagnosis, prognosis, and biology.
Authors
- Yong Wan (ORCID: https://orcid.org/0000-0002-3745-3447)
- Shuo Huang (ORCID: https://orcid.org/0000-0001-9495-4339)
- Rodrigo Ledesma‐Amaro (ORCID: https://orcid.org/0000-0003-2631-5898)
- Xi Zhang (ORCID: https://orcid.org/0000-0002-4638-3997)
- Yalin Zhang (ORCID: https://orcid.org/0000-0001-8955-8139)
- Haowei He
- Shuai Zhou (ORCID: https://orcid.org/0000-0001-8143-0413)
- Tao Yang (ORCID: https://orcid.org/0000-0002-7698-3440)
- Shanrong Liu
- Haimei Mao
- Qingshan Wei
- Chuanbin Mao
- D P Yin
- Xinyi Wan
- Anyi Li
- Yi Shen
- Yangdao Wei
- Xiangpeng Li
- Rui Wang
- Fengge Song
- Buhua Wang
- Ruijie Deng
- Zhiqing Yang
Institutions
- Florida State University (US)
- North Carolina State University (US)
- Central South University (CN)
- Army Medical University (CN)
- Chinese University of Hong Kong (HK)
- Second Military Medical University (CN)
- Hainan University (CN)
- Sichuan University (CN)
- Haikou City People's Hospital (CN)
- Changhai Hospital (CN)
- Hainan Center for Disease Control & Prevention (CN)
- Guangdong Testing Institute for Product Quality Supervision (CN)
- Imperial College London (GB)
- Hainan Medical University (CN)
Publication Details
- Journal
- Nature Communications
- Published
- 2026-02-12
- DOI
- https://doi.org/10.1038/s41467-026-69181-x
- Primary Topic
- MicroRNA in disease regulation
- Type
- article
- Field-Weighted Citation Impact
- 0.00