Cas12a-assisted split crRNA complex for analysis and detection of diverse entities

Nov 20, 2025Nucleic acids research

Using Cas12a with split guide RNA to detect and analyze different targets

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Abstract

A novel detection platform, , successfully identifies both nucleic acid and non-nucleic acid targets with high sensitivity and specificity.

  • The Cas12a-crRNA system is capable of recognizing specific DNA sequences and can cleave single-stranded DNA.
  • Key parameters for activating a split crRNA system were systematically explored, leading to established design guidelines.
  • CASCADE was validated for its effectiveness using microRNA, confirming its sensitivity and ability to distinguish between mismatched sequences.
  • Detection of non-nucleic acid targets, including tobramycin, kanamycin, biotin, and tetracycline repressor protein, was successfully demonstrated.
  • The integration of a lateral flow assay enhances the portability and ease of use of the CASCADE platform for on-site detection.

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Key numbers

100 pM
Sensitivity for let-7d
Detection limit for let-7d using .
0.175 μM
Detection limit for tobramycin
Lowest concentration of tobramycin detectable by .
0.2 μM
Detection limit for kanamycin
Lowest concentration of kanamycin detectable by .

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