Structural foundations of optogenetics
Abstract/Contents
- Abstract
- The brain is a remarkably complex structure, composed of hundreds of neurons in simple organisms and up to hundreds of billions of neurons in large mammals. The recent advent of optically-modulated, molecular tools for neuroscience ('optogenetics') has allowed unprecedented access to simultaneously modulate and observe the activity of hundreds of genetically-defined neurons with millisecond resolution. However, while optogenetics has enabled rapid advances in neuroscience, this powerful toolset remains constrained by a limited mechanistic understanding of light-gated molecules, including channelrhodopsins (ChRs). Here, I describe my efforts to understand structural and dynamical mechanisms of ChRs, using three complimentary approaches. First, to extend the available high-resolution ChR structural insights, I employed X-ray crystallography to determine the structures of natural and designed anion-conducting ChRs (ACRs). Next, I used a combination of these atomic-resolution structures, molecular dynamics (MD) computational simulation, and in vitro electrophysiology to assess functional dynamics of ACRs, leading to the identification of a variant with improved channel-closing kinetics. Finally, I used structure-guided genome mining, whole-cell patch clamp electrophysiology, and two-photon imaging to identify and characterize a new red-shifted excitatory channelrhodopsin with large photocurrents and high light sensitivity. Taken together, this work provides a framework for the engineering and discovery of better optogenetic tools and lays a foundation for future studies of channelrhodopsin biology
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2020; ©2020 |
| Publication date | 2020; 2020 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Kim, Yoon Seok |
|---|---|
| Degree supervisor | Deisseroth, Karl |
| Degree supervisor | Kobilka, Brian K |
| Thesis advisor | Deisseroth, Karl |
| Thesis advisor | Kobilka, Brian K |
| Thesis advisor | Lin, Michael Z |
| Degree committee member | Lin, Michael Z |
| Associated with | Stanford University, Department of Bioengineering. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Yoon Seok Kim |
|---|---|
| Note | Submitted to the Department of Bioengineering |
| Thesis | Thesis Ph.D. Stanford University 2020 |
| Location | electronic resource |
Access conditions
- Copyright
- © 2020 by Yoon Seok Kim
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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