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A switchable Cas12a enabling CRISPR-based direct histone deacetylase activity detection.
- Source :
-
Biosensors & Bioelectronics . Oct2022, Vol. 213, pN.PAG-N.PAG. 1p. - Publication Year :
- 2022
-
Abstract
- The efficient and robust signal reporting ability of CRISPR-Cas system exhibits huge value in biosensing, but its applicability for non-nucleic acid analyte detection relies on the coupling of additional recognition modules. To address this limitation, we described a switchable Cas12a and exploited it for CRISPR-based direct analysis of histone deacetylase (HDAC) activity. Starting from the acetylation-mediated inactivation of Cas12a by anti-CRISPR protein AcrVA5, we demonstrated that the acetyl-inactivated Cas12a could be reversibly activated by HDAC-mediated deacetylation based on computational simulations (e.g., deep learning and protein-protein docking analysis) and experimental verifications. By leveraging this switchable Cas12a for both target sensing and signal amplification, we established a sensitive one-pot assay capable of detecting deacetylase sirtuin-1 with sub-nanomolar sensitivity, which is 50 times lower than the standard two-step peptide-based assay. The versability of this assay was validated by the sensitive assessment of cellular HDAC activities in different cell lines with good accuracy, making it a valuable tool for biochemical studies and clinical diagnostics. [ABSTRACT FROM AUTHOR]
- Subjects :
- *HISTONE deacetylase
*DEACETYLATION
*DEEP learning
*SIRTUINS
*CRISPRS
*CELL lines
Subjects
Details
- Language :
- English
- ISSN :
- 09565663
- Volume :
- 213
- Database :
- Academic Search Index
- Journal :
- Biosensors & Bioelectronics
- Publication Type :
- Academic Journal
- Accession number :
- 157762901
- Full Text :
- https://doi.org/10.1016/j.bios.2022.114468