BioTechniques (Apr 2023)

Colorimetric miRNA detection based on self-primer-initiated CRISPR-Cas12a-assisted amplification

  • Ying Kang,
  • Jingjing Zhang,
  • Lijuan Zhao,
  • Hongli Yan

DOI
https://doi.org/10.2144/btn-2023-0008
Journal volume & issue
Vol. 74, no. 4
pp. 172 – 178

Abstract

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miRNAs alter significantly throughout pregnancy to support the development of the fetus. However, sensitive detection of miRNA remains a challenge. Herein, a reliable miRNA detection approach integrating self-assembly-triggered signal amplification and CRISPR-Cas12a-system cleavage-based color generation is described. The colorimetric approach contains three signal amplification processes. The first signal amplification is formed by the released miRNA in a chain extension process. The produced sequence that is similar to the target miRNA initiates the second signal recycle. Finally, CRISPR-Cas12a-based transcleavage on linker sequences induces the third signal amplification. The method exhibits high sensitivity and a low limit of detection of 254 aM, showing promising prospects in disease diagnosis.

Keywords