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Issue 83, 2020
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A method for the quantitative detection of Cas12a ribonucleoproteins

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Abstract

Cas12a ribonucleoprotein (RNP) is an RNA-guided CRISPR-associated nuclease used widely for genome editing and molecular diagnostics. Conventional detection methods rely on adopting antibody-based reagents that are expensive and lack scalability, and, moreover, only detect Cas12 enzyme rather than RNP, which is the true effector. Here, we describe a method for the rapid and quantitative detection of the effective Cas12a RNPs by the combined use of anti-CRISPR protein AcrVA1 and stem-loop RT-qPCR, achieving a limit of detection (LOD) of 1 fM in reaction buffer and 0.1 pM under biologically representative conditions.

Graphical abstract: A method for the quantitative detection of Cas12a ribonucleoproteins

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Supplementary files

Article information


Submitted
09 Jun 2020
Accepted
09 Sep 2020
First published
09 Sep 2020

Chem. Commun., 2020,56, 12616-12619
Article type
Communication

A method for the quantitative detection of Cas12a ribonucleoproteins

J. Qiao, S. Lin, W. Sun, L. Ma and Y. Liu, Chem. Commun., 2020, 56, 12616
DOI: 10.1039/D0CC04019C

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