Journal
ACS SYNTHETIC BIOLOGY
Volume 10, Issue 12, Pages 3600-3603Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.1c00455
Keywords
CRISPR; genome editing; biosensor; detection; transient gene expression
Categories
Funding
- Genomic Science Program of the U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research (BER), Secure Ecosystems Engineering and Design research program in the Secure Biosystems Design Scientific Focus Area (SFA)
- U.S. Department of Energy [DE-AC05-00OR22725]
- National Science Foundation Plant Genome Research Program [IOS-1758745, IOS-2029889]
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CRISPR/Cas technology has emerged as the most reliable system for genome engineering in various species, but concerns about potential unintended DNA changes from CRISPR gene editing are increasing. Developing a system that can detect and report the presence of active CRISPR/Cas tools in biological systems is essential. Researchers have developed four real-time detection systems that can effectively detect the activities of CRISPR/Cas9 nuclease, base editing, prime editing, and CRISPRa in plants.
CRISPR/Cas has recently emerged as the most reliable system for genome engineering in various species. However, concerns about risks associated with the CRISPR/Cas technology are increasing on potential unintended DNA changes that might accidentally arise from CRISPR gene editing. Developing a system that can detect and report the presence of active CRISPR/Cas tools in biological systems is therefore very necessary. Here, we developed four real-time detection systems that can spontaneously indicate the presence of active CRISPR-Cas tools for genome editing and gene regulation including CRISPR/Cas9 nuclease, base editing, prime editing, and CRISPRa in plants. Using the fluorescence-based molecular biosensors, we demonstrated that the activities of CRISPR/Cas9 nuclease, base editing, prime editing, and CRISPRa can be effectively detected in transient expression via protoplast transformation and leaf infiltration (in Arabidopsis, poplar, and tobacco) and stable transformation in Arabidopsis.
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