A5069998775- RNA and protein synthesis mechanisms
- CRISPR and Genetic Engineering
- Advanced biosensing and bioanalysis techniques
- RNA regulation and disease
University of Florida
2023
Abstract Cas12a, a CRISPR-associated protein complex, has an inherent ability to cleave DNA substrates and is utilized in diagnostic tools identify molecules. We demonstrate that multiple orthologs of Cas12a activate trans-cleavage the presence split activators. Specifically, PAM-distal region crRNA recognizes RNA targets provided PAM-proximal seed target. Our method, Split Activator for Highly Accessible Analysis (SAHARA), detects picomolar concentrations without sample amplification,...
Abstract CRISPR is a prominent bioengineering tool and the type V CRISPR-associated protein complex, Cas12a, widely used in diagnostic platforms due to its innate ability cleave DNA substrates. Here we demonstrate that Cas12a can also be programmed directly detect RNA substrates without need for reverse transcription or strand displacement. We discovered while PAM-proximal “seed” region of crRNA exclusively recognizes initiating trans-cleavage, PAM-distal 3’-end tolerate both Utilizing this...
CRISPR is a prominent bioengineering tool and the type V CRISPR-associated protein complex, Cas12a, widely used in diagnostic platforms due to its innate ability cleave DNA substrates. Here we demonstrate that Cas12a can also be programmed directly detect RNA substrates without need for reverse transcription or strand displacement. We discovered while PAM-proximal "seed" region of crRNA exclusively recognizes initiating trans- cleavage, PAM-distal 3'-end tolerate both Utilizing this...