BCCA Terry Fox Lab Special Seminar - Dr. David J.H.F. Knapp
The advent of the CRISPR/Cas9 system has opened the floodgates for experiments which require precise gene editing. This system relies on two components, a guide RNA which determines gene specificity, and the protein Cas9 which performs effector functions. Guide RNA are generally produced using constitutive Pol-III promoters, as processing is required for proper function when produced from a Pol-II promoter. Several reports suggest that t-RNA can be used to process guides out of longer Pol-II transcripts. We have found, however, that diverse t-RNA from fly to human, have constitutive Pol-III promoter activity thus disallowing their use in inducible systems. In order to correct this deficiency we have performed a combination of curated mutations, and high throughput mutational screening to independently identify the features required for the promoter activity and processing ability of human t-RNA.
Host: Dr. Connie Eaves
Distinguished Scientist, Terry Fox Lab
|Knapp D Seminar Notice 060817.pdf||94.16 KB|