Application and characterization of optogenetic systems based on cryptochrome 2/CIB1
Abstract/Contents
- Abstract
- Various cellular processes are dependent on the regulation of protein activities. In particular, some cellular processes, including migration, division and differentiation, require sophisticated coordination of protein activities in space and time. Therefore, a method capable of perturbing protein activities with precise spatial and temporal control is indispensable for understanding cellular behaviors. However, conventional means, such as genetic or pharmacological perturbation, are either relatively slow to implement, difficult to design, or spatiotemporally uncontrollable. Approaches using inducible chemical genetic enable the control of protein activities by using chemical inducers to trigger engineered proteins. However, these approaches suffer from irreversibility and cannot achieve spatial control. The emerging area of optogenetic actuators provides opportunities to temporally and spatially regulate protein activities. Several pairs of light-induced dimerization proteins have been developed, including the light-oxygen-voltage domain, phytochrome B and cryptochrome 2 (CRY2). CRY2 and its binding partner CIB1 are the optogenetic dimerizers utilized in my research due to their rapidness, reversibility and no need for exogenous cofactor. In my thesis, I develop the optogenetic strategies to optically manipulate two processes: signaling pathway and organelle activity, both of which are determined by protein activities. The light-induced CRY2/CIB1 interaction is complicated by the light-induced CRY2 homo-oligomerization. Therefore, in this thesis, I also study the dual characteristics of CRY2 homo-oligomerization and hetero-dimerization. This thesis consists of three chapters. In the first chapter, I demonstrate the strategies designed to optically activate the RAF/ERK and AKT/FOXO signaling pathways with temporal control, which can help to dissect and resolve the signaling pathways in a quantitative manner. Next, I construct an optogenetic method that exploits light to manipulate organelle distribution and reshaping with reversibility and subcellular spatial precision. This method will be useful to establish a direct linkage between organelle distribution/shaping and cellular functions. Finally, I characterize the CRY2 homo-oligomerization and its interplay with CRY2/CIB1 hetero-dimerization. The results can serve as a guide to the usage of CRY2/CIB1 system.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2016 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Duan, Liting | |
|---|---|---|
| Associated with | Stanford University, Department of Chemistry. | |
| Primary advisor | Cui, Bianxiao | |
| Thesis advisor | Cui, Bianxiao | |
| Thesis advisor | Dai, Hongjie, 1966- | |
| Thesis advisor | Fayer, Michael D | |
| Advisor | Dai, Hongjie, 1966- | |
| Advisor | Fayer, Michael D |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Liting Duan. |
|---|---|
| Note | Submitted to the Department of Chemistry. |
| Thesis | Thesis (Ph.D.)--Stanford University, 2016. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2016 by Liting Duan
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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