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Engineering Proximity Labeling Techn...

Cheah, Joleen,

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Engineering Proximity Labeling Techniques to Map Translocating Proteomes and Rna Reporters to Surveil Cell States /

Record Type:	Electronic resources : Monograph/item
Title/Author:	Engineering Proximity Labeling Techniques to Map Translocating Proteomes and Rna Reporters to Surveil Cell States // Joleen Cheah.
Author:	Cheah, Joleen,
Description:	1 electronic resource (267 pages)
Notes:	Source: Dissertations Abstracts International, Volume: 86-05, Section: B.
Contained By:	Dissertations Abstracts International86-05B.
Subject:	Cancer. -
Online resource:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=31643292
ISBN:	9798346383949

Engineering Proximity Labeling Techniques to Map Translocating Proteomes and Rna Reporters to Surveil Cell States /
Cheah, Joleen,

Engineering Proximity Labeling Techniques to Map Translocating Proteomes and Rna Reporters to Surveil Cell States /Joleen Cheah. - 1 electronic resource (267 pages)

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

Proximity 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.

English

ISBN: 9798346383949Subjects--Topical Terms:

634186
Cancer.

Engineering Proximity Labeling Techniques to Map Translocating Proteomes and Rna Reporters to Surveil Cell States /
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520 $a Proximity 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.
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650 4 $a Proteomics. $3 600542
650 4 $a Analytical chemistry. $3 3168300
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