RT Journal Article
SR Electronic
T1 Engineering strategy and vector library for the rapid generation of modular light-controlled protein-protein interactions
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 583369
DO 10.1101/583369
A1 Alexandra-Madelaine Tichy
A1 Elliot J. Gerrard
A1 Julien M.D. Legrand
A1 Robin M. Hobbs
A1 Harald Janovjak
YR 2019
UL http://biorxiv.org/content/early/2019/03/20/583369.abstract
AB Optogenetics enables the spatio-temporally precise control of cell and animal behaviour. Many optogenetic tools are driven by light-controlled protein-protein-interactions (PPIs) that are repurposed from natural light-sensitive domains (LSDs). Applying light-controlled PPI to new target proteins is challenging because it is difficult to predict whether one the many available LSDs will yield robust light regulation. As a consequence, fusion protein libraries need to be prepared and tested, but methods and platforms to facilitate this process are currently not available. Here, we developed a genetic engineering strategy and vector library for the rapid generation of light-controlled PPIs. The strategy permits fusing a target protein to LSDs efficiently and in two orientations. The public and expandable library contains 29 vectors with blue, green or red light-responsive LSDs many of which have been previously applied ex vivo and in vivo. We demonstrate the versatility of the approach and the necessity for sampling LSDs by generating light-activated caspase-9 (casp9) enzymes. Collectively, this work provides a new resource for optical regulation of a broad range of target proteins in cell and developmental biology.