Smart Solutions in drug discovery
Poster Nov 13, 2009
Ian Kalinka, Lynne Smith, Elizabeth Price, Angela Marenghi, J.M.Kendall, Ray Ismai
Here we present data on the development of a large scale production process for the provision of two different cryopreserved transiently transfected cell lines. Included is identification of a suitable transfection reagent that is efficient, non-toxic and cost effective. This process has been validated using these two cell lines and two different cell-based assays.
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