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From Single-Cell to Whole-Body: Developing a Molecular Neuroscience Toolkit

Citation

Flytzanis, Nicholas C. (2019) From Single-Cell to Whole-Body: Developing a Molecular Neuroscience Toolkit. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/S28C-DJ17. https://resolver.caltech.edu/CaltechTHESIS:10122018-165154970

Abstract

Throughout my Ph.D. I have worked on technology development, at first to answer basic scientific questions and eventually for therapeutic applications. This technology development applied to a variety of fields, from neuroscience to development to gene therapy, and acted upon biological systems in a wide range of scale, from the single-cell monitoring to organism-wide gene-transfer. My graduate research began with the engineering of microbial rhodopsin spectral properties and fluorescence. By making use of their ability to absorb light and emit fluorescence in a voltage-dependent manner, I aimed to interrogate neuronal activity during behavior at the single-cell level. That line of research ended with publication of the voltage-sensor Archer, which I used to track activity of a single cell in vivo in awake, behaving worms. I then shifted from tracking activity at the single cell level, to visualizing entire organisms, by developing clearing techniques that enable a high-resolution, three-dimensional analysis of a diverse range of tissues. I began by optimizing tissue-clearing parameters for various tissue types and a wide variety of experimental needs. I then took that knowledge and applied it to visualizing and tracking the developing neural crest in cleared, whole-mount chicken embryos, discovering some unexpected derivates. Finally, I became interested not only in visualizing entire organisms, but in developing technologies to facilitate gene transfer throughout the body. The rapidly growing field of gene therapy is in constant need of new tools that target specific tissues, avoiding off-target effects. The end of my Ph.D. has been spent engineering viruses that can be delivered body-wide, but target only specific areas of therapeutic interest, like the brain and lungs.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:Tool development, Neuroscience, Optogenetics, Voltage sensors, Tissue clearing, Directed evolution, Viral engineering, AAV
Degree Grantor:California Institute of Technology
Division:Biology and Biological Engineering
Major Option:Biology
Awards:Demetriades-Tsafka-Kokkalis Prize in Entrepreneurship or Related Fields, 2019.
Thesis Availability:Public (worldwide access)
Research Advisor(s):
  • Gradinaru, Viviana
Thesis Committee:
  • Anderson, David J. (chair)
  • Arnold, Frances H.
  • Shapiro, Mikhail G.
  • Gradinaru, Viviana
Defense Date:10 September 2018
Non-Caltech Author Email:nickflytzanis (AT) gmail.com
Record Number:CaltechTHESIS:10122018-165154970
Persistent URL:https://resolver.caltech.edu/CaltechTHESIS:10122018-165154970
DOI:10.7907/S28C-DJ17
Related URLs:
URLURL TypeDescription
https://doi.org/10.1038/529469aDOIArticle adapted for Chapter 2
https://doi.org/10.1016/j.jmb.2014.06.015DOIArticle adapted for Chapter 2.2
https://doi.org/10.1073/pnas.1413987111DOIArticle adapted for Chapter 2.3
https://doi.org/10.1038/ncomms5894DOIArticle adapted for Chapter 2.4
https://doi.org/10.1038/nprot.2015.122DOIArticle adapted for Chapter 3.2
ORCID:
AuthorORCID
Flytzanis, Nicholas C.0000-0002-7921-9392
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:11230
Collection:CaltechTHESIS
Deposited By: Nicholas Flytzanis
Deposited On:22 Oct 2018 21:16
Last Modified:22 Oct 2019 21:25

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