High-Throughput Multiplexed Inflammation Assay Using Primary Cells
Poster May 11, 2010
Oksana Sirenko, Yen-Wen Chen, Jayne Hesley, Cathy Olsen, H. Roger Tang, Evan F. Cromwell*
An important event in the inflammation response is the expression of cell-surface antigens that facilitate binding of immune cells to blood vessels. The ability to monitor up-regulation of these molecules, such as VCAM, E-selectin, and HLADR on endothelial cells provides an important physiological read-out for cell-based models of inflammation.
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