GPVI-RBL-2H3-NFAT-Re Luciferase Cell Line: a New Cell Based System for Studying Collagen Receptor Activation
Poster Jun 08, 2005
Marcella De Silvestris, Claudia Caserini, Maria Grazia Giribaldi, Silvia Bovolenta and Lia Scarabottolo
The response of platelets to vessel wall injury is a primary event in arterial thrombosis. Platelets respond to vessel wall injury via activation of the Collagen surface receptor Glycoprotein VI (GPVI), which accelerates the thrombotic response through release of granule contents and activation of platelet integrins.
GPVI, which represents a relevant antithrombotic pharmacological target, is a platelet membrane protein constitutively associated with the Fc Receptor y chain (FcR y). Upon activation by the specifi c ligand Collagen, GPVI induces a kinase cascade, resulting in the stimulation of PLC-y2 which fi nally leads to cytosolic Ca++ increase (Figure 1).
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