NFAT THESIS PROJECT
For my PhD thesis project, I adapted the optogenetic tool CLASP (Controllable Light Activated Shuttling and
Plasma membrane sequestration) from yeast to mammalian cells. This figure illustrates the mechanism of CLASP.
CLASP utilizes two asLOV2 domains, blue light sensitive proteins derived from Avena Sativa, to sequester protein cargo at the plasma membrane. In response to blue light, the asLOV2 domains release the cargo and expose a nuclear localization signal.
Below is an overview of the endogenous NFAT nuclear shuttling mechanism.
And lastly, below is a high level overview of how cells use a finite number of signals to produce a wide range of physiological responses, and how we use a combination of optogenetics and differential gene expression analyses to stratify the significance of nuclear translocation alone on transcription.
