Journal
NATURE CHEMICAL BIOLOGY
Volume 12, Issue 6, Pages 425-U78Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/NCHEMBIO.2063
Keywords
-
Categories
Funding
- National Institutes of Health [GM100225]
- McKnight Endowment Fund for Neuroscience (Technological Innovations in Neuroscience Award)
Ask authors/readers for more resources
Arabidopsis thaliana cryptochrome 2 (AtCRY2), a light-sensitive photosensory protein, was previously adapted for use in controlling protein-protein interactions through light-dependent binding to a partner protein, CIB1. While the existing CRY2-CIB dimerization system has been used extensively for optogenetic applications, some limitations exist. Here, we set out to optimize function of the CRY2-CIB system by identifying versions of CRY2-CIB that are smaller, show reduced dark interaction, and maintain longer or shorter signaling states in response to a pulse of light. We describe minimal functional CRY2 and CIB1 domains maintaining light-dependent interaction and new signaling mutations affecting AtCRY2 photocycle kinetics. The latter work implicates an alpha 13-alpha 14 turn motif within plant CRYs whose perturbation alters signaling-state lifetime. Using a long-lived L348F photocycle mutant, we engineered a second-generation photoactivatable Cre recombinase, PA-Cre2.0, that shows fivefold improved dynamic range, allowing robust recombination following exposure to a single, brief pulse of light.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available