Trial research of fibrous dysplasia model transplanted with GNAS1 mutant cells

为了用His取代Gs中的Arg201，我们制备了GNAS1突变基因（R201H），以A代替G，从而导致人纤维性骨发育不良（FD）。为了将R201H稳定地转移到培养细胞中，我们构建了表达R201H的慢病毒载体，并利用它转染C57BL/6小鼠骨髓基质细胞。我们现在正在研究表达r201h的细胞的增殖和分化，并准备通过原位移植的方法制作小鼠模型。此外，我们研究了高度精确的基因测试方法，我们可以将其与其他类似的骨病变区分开来。因此，与传统的限制性片段长度多态性PCR相比，使用PNA探针和焦磷酸测序方法的实时PCR更有效。

To replacement Arg201 with His in the Gs, we produced mutated GNAS1 gene (R201H) replacing with A instead of G of the human GNAS1 gene, which causes human fibrous dysplasia of bone (FD). To stably transfer the R201H into cultured cells, we constructed a lentiviral vector expressing R201H, and used it to transduce C57BL/6 mice-derived marrow stromal cells. We are now elucidating the cell proliferation and differentiation of the R201H-expressing cells and are going to make a mouse model by the orthotopic transplant of the transduced cells. In addition, we studied the method of highly precise genetic tests that we could distinguish it from other similar bone lesions. As a result, the real-time PCR using PNA probe and pyrosequencing method were more effective, compared to conventional PCR-restriction fragment length polymorphism.