Determination of Cell Colony Formation in a High Content Screening Assay
Poster Mar 13, 2006
David Pole and Wayne Bowen
AbstractHere, laser scanning microplate cytometry has been used to provide an automated high content readout of the effects of cytostatic agents on colony formation. This approach determines colony number through the application of a spherical volume algorithm. This permits the differentiation of cytostatic effects where the number of colonies and size remains constant and cytotoxic effects where the size and number may be reduced.
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