VVD Engineering And CRY2Application Of Optogenetic Tool

Optogenetics combines recombinant DNA technology and optic technology, It has been widely used for tracing target protein in living cell as well as selectively controlling preciseneural activity patterns within subtypes of cells of the brain for in depth study of neuroscience research. Recentlyoptogenetics has been extended to the study of signal transduction and even be explored for clinical application. Furtherdevelopment of optogenetics will no doubt feasible the study of synthetic physiology. Optogenetics wasselected by the journal of Nature Methods as method of the year2010.Tools for optical control of proteins offer an unprecedented level of spatiotemporal controlover biological processes, adding a new layer of experimental opportunity. While use of light-activated cation channels and anion pumps has already revolutionized neurobiology, an emerging class of more general optogenetic tools may have similar transformative effects. These tools consist of light-dependent protein interaction modules that allow control of target protein interactions and localization with light. Such tools are modular and can be applied to regulate a wide variety of biological activities. There are the different properties of light-induced dimerization systems, based on plant phytochromes, cryptochromes, and light-oxygen-voltage domain proteins, exploring advantages and limitations of the different systems and practical considerations related to their use. Potential applications of these tools within the neurobiology field, including light control of various signaling pathways, neuronal activity, and DNA recombination and transcription, and so on.optogenetic tool has widely apply in the future, it remain has some unclear region in the organelles, heterodimer tool still not enough, the regulative wavelength is not suit for using. Aiming at the problem, these problem become focus in the study.1light-induce ion channel. Arabidopsis thaliana cryptochrome2(CRY2) for rapid and reversible protein oligomerization was response to blue light. STIM1is universally distributed in ER membrane at rest and activates the CRAC channel by aggregating underneath the plasma membrane (PM) and co-clustering with Orail at ER-PM junctions. CRAC have spatiotemporal precise and reversible control under CRY2oligomerization2Engeerniering of light control protein-Vivid(VVD), VVD form homodimer under blue light, VVD has more widely use in the future to manipulate biological processes in living system with minimal perturbation while investigating and predicating it structure to mutant it amino acid becoming a heterodimer toolResult show that CRY2could not oligomerization as cytoplasm needing to optimize further, after engineering, VVD become a good pair----52K55F with55L56D which only have15%homodimer, but130%heterodimer.