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).


