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<p>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 (<Emphasis Type="Underline">P</Emphasis>NA) mediated C<Emphasis Type="Underline">RI</Emphasis>SPR/<Emphasis Type="Underline">C</Emphasis>as13a syst<Emphasis Type="Underline">e</Emphasis>m (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.</p>

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PRICE: direct and robust detection of microRNAs at single-nucleotide resolution

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年2月12日

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  • Buhua Wang,
  • Shuai Zhou,
  • Xi Zhang,
  • Rui Wang,
  • Shuo Huang,
  • Anyi Li,
  • Haowei He,
  • Yingai Zhang,
  • Yangdao Wei,
  • Zhiqing Yang,
  • Fengge Song,
  • Xiangpeng Li,
  • Xinyi Wan,
  • Yi Shen,
  • Chunxin Ma,
  • Haimei Mao,
  • Rodrigo Ledesma-Amaro,
  • Dapeng Yin,
  • Qingshan Wei,
  • Ruijie Deng,
  • Tao Yang,
  • Shanrong Liu,
  • Yi Wan,
  • Chuanbin Mao

摘要

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.