On relationships between cellular spatiotemporal dynamics and behavior
Abstract/Contents
- Abstract
- Biological cells are intricately organized on many length and timescales. Here, we investigate the organization of DNA and its transcription in the nucleus of human cells (Chapter 2), the translation and organization of proteins in the cytoplasm (Chapter 3), and gradual changes in the chromatin during differentiation (Chapter 4). A common thread through all of this work is the notion that precision measurement tools, especially advanced optical techniques in concert with new probes, are essential for discovering new biology. Chapter 2 explores interactions among chromatin, transcription factors, and potential 'genome organizers'. Special AT-rich protein 1 (Satb1) is a single protein important for proper mammalian development and immune system, that integrates all three functions, a transcription factor crucial for proper mammalian development, the immune system, and a potential genome organizer sits at the epicenter of these topics. Genomics, live cell single molecule imaging, and \textit{in vitro} DNA-protein binding assays help paint a detailed and dynamic portrait of Satb1 chromatin interactions and provides compelling evidence of Satb1 as a pioneer factor. Chapter 3 covers the development of a live cell imaging platform for visualization of endogenous translation forms using a CRISPR/Cas9 based genome editing strategy. This platform was employed to study endogenous paxillin isoforms in focal adhesions. Using this visualization platform in concert with high spatial-resolution imaging, we discovered preliminary evidence for nanoscale clustering and organization of paxillin isoforms in focal adhesions. Chapter 4 probes chromatin dynamics in differentiating human embryonic stem cells (hESCs). We established a human embryonic stem cell line for live cell single molecule tracking of nucleosomes harboring fluorescent H2B molecules, allowing for dynamic tracking of chromatin motion at single molecule resolution in pre and post differentiated stem cells. Together, the work demonstrates the use of novel imaging tools for understanding how cells organize two of their major components, chromatin in the nucleus and focal adhesions in the cytoplasm
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 | Reddick, Michael Princton |
|---|---|
| Degree supervisor | Liphardt, Jan |
| Degree supervisor | Spakowitz, Andrew James |
| Thesis advisor | Liphardt, Jan |
| Thesis advisor | Spakowitz, Andrew James |
| Thesis advisor | Greenleaf, William James |
| Degree committee member | Greenleaf, William James |
| Associated with | Stanford University, Department of Chemical Engineering. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Michael Reddick |
|---|---|
| Note | Submitted to the Department of Chemical Engineering |
| Thesis | Thesis Ph.D. Stanford University 2020 |
| Location | electronic resource |
Access conditions
- Copyright
- © 2020 by Michael Princton Reddick
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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