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Runx2 expression analysis in osteoblast patho-physiology

Student name: Ms Komal Kumari
Guide: Dr Chaithanya Madhurantakam
Year of completion: 2018
Host Organisation: National Institute of Immunology, New Delhi
Supervisor (Host Organisation): Dr Vijay K. Yadav
Abstract: Bone is living, growing tissue. It is made mostly of collagen, a protein that provides a soft framework, and calcium phosphate, a mineral that adds strength and hardens the framework. This combination of collagen and calcium makes bone both flexible and strong, which in turn helps bone to withstand stress. More than 99 percent of the body’s calcium is contained in the bones and teeth. The remaining 1 percent is found in the blood.

Throughout one’s lifetime, old bone is removed (resorption, known as osteoclast) and new bone is added to the skeleton (formation, known as osteoblast). Osteoblast cells are derived from mesenchymal cells . And the transcription factor, responsible to differentiate osteoblast lineage is Cbfa1/ Runx2. It is a master regulator for osteoblast differentiation.

To analyse the Runx2 promoter activity, amplified 2kb, 5’ flanking region of Runx2 distal or P1 promoter. This 2 kb promoter region is cloned in luciferase vector to generate luciferase reporter construct. A luciferase reporter construct is analyzed in mouse MC3T3-E1, pre-osteoblast cell line which has a potency to undergo differentiation and maturation. The activity of runx2 promoter is examined under hypertonic condition through dual luciferase reporter assay.

This project facilitated us with a tool to study the effect of different physiological condition like hypoxia, hypertonicity on growth and development.