Real Time Gene Expression Analysis In Live Cells

Investigator: Hetal Desai

Stem cell differentiation is a complex process that can be difficult to detect in early stages. Lineage-specific mRNA, however, are transcribed at different stages, and a subset could serve as early biomarkers of differentiation. One method to detect lineage-specific mRNA is to develop a robust, transiently applied live cell imaging assay. To this end, molecular beacons, fluorescently functionalized nucleic acid hybridization probes, are an excellent visualization tool
capable of imaging the mRNA of various genes in live cells without altering gene function. We have developed novel molecular beacons for three osteogenic lineage-specific genes: alkaline phosphatase (ALP), collagen 1 (COL1A1) and osteocalcin (BGLAP) and tested them in both MG63 (human osteosarcoma) cells and differentiating adipose-derived stem cells (ASCs).