자료유형  

 학위논문

Control Number  

0017164802

International Standard Book Number  

9798346383949

Dewey Decimal Classification Number  

616

Main Entry-Personal Name  

Cheah, Joleen.

Publication, Distribution, etc. (Imprint  

[S.l.] : Stanford University., 2024

Publication, Distribution, etc. (Imprint  

Ann Arbor : ProQuest Dissertations & Theses, 2024

Physical Description  

267 p.

General Note  

Source: Dissertations Abstracts International, Volume: 86-05, Section: B.

General Note  

Advisor: Ting, Alice.

Dissertation Note  

Thesis (Ph.D.)--Stanford University, 2024.

Summary, Etc.  

요약Proximity labeling has been a revolutionary tool that has enabled many researchers to explore the protein space surrounding their areas of interest. The first half of my thesis, Chapter 1-3, focuses on my efforts to expand the capabilities of previously available proximity labeling tools. Chapter 1 focuses on the development of a light gated TurboID, LOV-Turbo, to provide this temporal specificity in cases where biotin is readily abundant, such as in neuron cultures and mice brains. In addition to temporal specificity, light-gating also enables spatial specificity to combat instances when TurboID cannot be cleanly targeted. Chapter 2 addresses the limitations of using proximity labeling enzymes and fractionation to study translocating proteomes due to impure fractions. Chapter 2 describes TransitID, a tandem labeling protocol with TurboID and APEX to identify proteins trafficking between the 2 enzyme's localization. Chapter 3 further explores LOV-Turbo's utility by improving labeling efficiencies achieved by activation via bioluminescence resonance energy transfer. This enables further spatial specification of labeling though targeting of the bioluminescence emitting enzyme, luciferase.In the second half of this thesis, I describe my work on developing RNA recorders to log cellular events in RNA to generate a timeline of the events. Chapter 4 details an RNA recorder that is collected from the exosome fraction, enabling continuous monitoring without harming the cells. Therefore, multiple readouts can be analyzed then assembled together in temporal order to generate a timeline of the recorded event. Two methods of encoding information into RNA are introduced: event-dependent editing of exosome targeted RNA and event-dependent export of RNA to exosomes In Chapter 5, I propose another alternative to recording cellular events in RNA to not only provide information on if the event occurred but also when. This was designed by having two editing modalities on the RNA, one to log the occurrence of the event, and one to log when the event occurred. Further development of these tools to increase their temporal sensitivity would enable high throughput tracking of cellular events over time.

Subject Added Entry-Topical Term  

Cancer.

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Communication.

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Mutation.

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Bioluminescence.

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Data processing.

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Fractionation.

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Yeast.

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Cell culture.

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Protein synthesis.

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Mass spectrometry.

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Cloning.

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Design.

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Engineering.

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Phenols.

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Cell growth.

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Polypeptides.

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Proteomics.

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Analytical chemistry.

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Bioinformatics.

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Cellular biology.

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Organic chemistry.

Added Entry-Corporate Name  

Stanford University.

Host Item Entry  

Dissertations Abstracts International. 86-05B.

Electronic Location and Access  

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Control Number  

joongbu:654506