Droplet-on-Demand Platform for Biochemical Screening & Drug Discovery
Poster Mar 06, 2015
L.D. van Vliet1*, F. Gielen1, A. Sinha2, B.T. Koprowski3, J.B. Edel4, X.Niu5, A.J. deMello3, F. Hollfelder1, & J. Motschman2
The method for generating specific sequences of droplets is previously described. Now several modes are demonstrated to enable a wider range of applications.
• Sequences of droplets can be generated from 24 wells.
• Droplets can be merged pairwise.
• Nanoliter droplets can be split into picoliter droplets.
• Our system enables combinatorial screening in nanolitre volumes and we have demonstrated its application to drug screening whether in vitro or in vivo.
• Encapsulated S. Cerevisiae cells showed good growth for all cell strains overnight.
• C. elegans were also encapsulated and were stable in droplets overnight.
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