Regulation of Smad6 Gene Expression using Catalytic RNA: Suppression of Smad6 Protein Expression by Hammerhead Ribozymes
The significance of bone morphogenetic protein activity in vivo is the induction of new bone. BMP?s were identified as proteins that regulate a wide range of functions, such as proliferation, differentiation and apoptosis of many different types of cells, such as osteoblasts, epithelial cells, neurons and immune cells. BMP signaling pathway is regulated by Smad proteins, which include Smad6 that has the ability to inhibit the BMP pathway. This is of particular of interest in gene therapy whereby regulation of Smad6 expression using catalytic RNA might be able to stimulate enhancement in bone formation. In this project, hammerhead ribozymes were designed to target specifically at Smad6. Following the transfection of the ribozymes into the cells, ribozyme 1 and 4 indicate a significant level of Smad6 suppression at the average of 59% and 52%, respectively, relative to the control cells. Results obtained from this project may provide insight for regulating Smad6 in the BMP signaling pathway. As well as, a new gene therapeutic approach for treatment in generating bone, bone healing, repairing soft tissue and stabilizing bony skeleton.