Author: Long T. Nguyen; Brianna M Smith; Piyush K. Jain
Title: Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection Document date: 2020_4_14
ID: n5kpvsvg_20
Snippet: In summary, we extended the 3'-and 5'-end of the crRNA and discovered an amplified transcleavage activity of LbCas12a when the 3'-end is extended with DNA or RNA. We applied this modified crRNA/LbCas12a system with the optimal conditions to detect PCA3 in simulated urine with high sensitivity. This ENHANCE technology enabled us to detect the DNA/RNA heteroduplex and methylated DNA with unprecedented sensitivity. We further employed this system to.....
Document: In summary, we extended the 3'-and 5'-end of the crRNA and discovered an amplified transcleavage activity of LbCas12a when the 3'-end is extended with DNA or RNA. We applied this modified crRNA/LbCas12a system with the optimal conditions to detect PCA3 in simulated urine with high sensitivity. This ENHANCE technology enabled us to detect the DNA/RNA heteroduplex and methylated DNA with unprecedented sensitivity. We further employed this system to test a range of target nucleic acids, including ssDNA, dsDNA, and RNA from HIV, HCV, and SARS-CoV-2 without the need for further optimization. These findings are a crucial step towards enhancing detection of nucleic acids and assisting in the diagnosis of various diseases. author/funder. All rights reserved. No reuse allowed without permission.
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