Expression and Purification of PI3 Kinase alpha and Development of a Luminometric
Poster Feb 15, 2008
Human class Ia PI3 kinase p110alpha catalytic and p85alpha regulatory subunits were expressed and purified. A luminometric ATP depletion assay to measure PI3K activity was established and optimized. Furthermore, the autophosphorylation status of PI3K alpha was checked and the IC50 value for the known PI3K inhibitor wortmannin was determined. The assay is homogenous and is potentially suitable for high throughput screening to identify PI3K inhibitors.
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