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Probing gene expression in single cells
Most gene expression methods report the average expression in a large cell population, losing information from individual cells. Author now show that fluorescent nucleic acid probes known as molecular beacons can be used to monitor simultaneously the expression of multiple genes in single live cells. The probe fluoresces when it binds to its target, causing a conformational change that separates a quencher and fluorophore attached to either of its ends. As a demonstration, the researchers inject molecular beacons with different fluorophores directly into single cells to monitor expression of genes encoding ß-actin and manganese superoxide dismutase. They measure the expression of the genes before and after exposing the cells to lipopolysaccharide, which stimulates expression of manganese superperoxide dismutase. By taking the ratio of the fluorescence to that of a reference probe, the researchers can compare gene expression from cell to cell. Author plans to use the technique to study fundamental questions in neuroscience and cancer.
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