Comparative Virtual and Experimental High-Throughput Screening for Glycogen Synthase Kinase-3b Inhibitors
Poster Feb 01, 2007
György M. Keser, Tímea Polgár, Andrea Baki and Györgyi I. Szendrei
Glycogen synthase kinase-3b (GSK-3b ) is a serine/threonine kinase that has recently emerged as a key target for neurodegenerative diseases and diabetes.
As an initial step of our lead discovery program we developed a virtual screen to discriminate known GSK-3b inhibitors and inactive compounds using FlexX, FlexX-Pharm and FlexE.
The maximal enrichment factor (EF=28) suggests that our protocol identifies potential GSK-3b inhibitors effectively from large compound collections. The effectiveness of our screening protocol was further investigated by a comparative experimental and virtual high-throughput screens performed for the same subset of our corporate library.
Enrichment factors, the significantly higher hit rate of virtual screening (12.9%) than that of the HTS (0.55%) and also the comparison of active clusters suggest that our virtual screening protocol is an effective tool in GSK-3b -based library focusing. Head-to-head comparison of true/false positives and negatives, revealed the two approaches to be complementary rather than competitive.
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