Team:SUSTech Shenzhen
ABSTRACT
Recently, gene express regulation techniques such as optogenetics and chemical genetics are commonly used in biology science, however, they all have their own limitation of drawbacks. Here we design a new way, audiogenetics, to precisely and nontoxicly regulate the gene expression in cells. It used a membrane mechanosensitive channel transient receptor potential channel 5 (TRPC5) and mammalian mechanosensitive Piezo1 channel to transform the audio wave energy as the input signal from extracellular into intracellular downstream signal. To make the TRPC5 more sensitive to mechanical stress, we use protein engineering by evolution to get a mutated TRPC5 with a high sensitive to sound waves. The downstream signal is calcium ion, which is the second messenger in cells and we use calcium indicator (R-GECO) to quantify the intracellular calcium by using live cell image. Calcium in cytosolic can regulate a series of phosphorylation and we know that it can induce the specific promoters (PNFAT) transgene expression. Finally, we use the GFP as the output signal to quantitatively analysis the regulatory ability of audiogenetics.
