Two secreted orthogonal bioluminescent reporters for non-destructive ultrasensitive assays
Poster May 11, 2010
George Tzertzinis1, Salvatore Russello1, Cathy Olsen2
Here we present a powerful combination of two secreted bioluminescent reporters, the luciferases from Gaussia princeps and Cypridina noctiluca. We demonstrate that the properties of these two reporters in terms of sensitivity, compatibility and use in non- destructive assays conducted on live cells or supernatants from different well formats makes them suitable for novel high throughput format applications.
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE