서브메뉴
검색
Engineering Proximity Labeling Techniques to Map Translocating Proteomes and Rna Reporters to Surveil Cell States
Engineering Proximity Labeling Techniques to Map Translocating Proteomes and Rna Reporters to Surveil Cell States
상세정보
- 자료유형
- 학위논문 서양
- 최종처리일시
- 20250211153048
- ISBN
- 9798346383949
- DDC
- 616
- 저자명
- Cheah, Joleen.
- 서명/저자
- Engineering Proximity Labeling Techniques to Map Translocating Proteomes and Rna Reporters to Surveil Cell States
- 발행사항
- [Sl] : Stanford University, 2024
- 발행사항
- Ann Arbor : ProQuest Dissertations & Theses, 2024
- 형태사항
- 267 p
- 주기사항
- Source: Dissertations Abstracts International, Volume: 86-05, Section: B.
- 주기사항
- Advisor: Ting, Alice.
- 학위논문주기
- Thesis (Ph.D.)--Stanford University, 2024.
- 초록/해제
- 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.
- 일반주제명
- Cancer
- 일반주제명
- Communication
- 일반주제명
- Mutation
- 일반주제명
- Bioluminescence
- 일반주제명
- Data processing
- 일반주제명
- Fractionation
- 일반주제명
- Yeast
- 일반주제명
- Cell culture
- 일반주제명
- Protein synthesis
- 일반주제명
- Mass spectrometry
- 일반주제명
- Cloning
- 일반주제명
- Design
- 일반주제명
- Engineering
- 일반주제명
- Phenols
- 일반주제명
- Cell growth
- 일반주제명
- Polypeptides
- 일반주제명
- Proteomics
- 일반주제명
- Analytical chemistry
- 일반주제명
- Bioinformatics
- 일반주제명
- Cellular biology
- 일반주제명
- Organic chemistry
- 기타저자
- Stanford University.
- 기본자료저록
- Dissertations Abstracts International. 86-05B.
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
008250123s2024 us c eng d■001000017164802
■00520250211153048
■006m o d
■007cr#unu||||||||
■020 ▼a9798346383949
■035 ▼a(MiAaPQ)AAI31643292
■035 ▼a(MiAaPQ)Stanforddd719tt2345
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a616
■1001 ▼aCheah, Joleen.
■24510▼aEngineering Proximity Labeling Techniques to Map Translocating Proteomes and Rna Reporters to Surveil Cell States
■260 ▼a[Sl]▼bStanford University▼c2024
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2024
■300 ▼a267 p
■500 ▼aSource: Dissertations Abstracts International, Volume: 86-05, Section: B.
■500 ▼aAdvisor: Ting, Alice.
■5021 ▼aThesis (Ph.D.)--Stanford University, 2024.
■520 ▼aProximity 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.
■590 ▼aSchool code: 0212.
■650 4▼aCancer
■650 4▼aCommunication
■650 4▼aMutation
■650 4▼aBioluminescence
■650 4▼aData processing
■650 4▼aFractionation
■650 4▼aYeast
■650 4▼aCell culture
■650 4▼aProtein synthesis
■650 4▼aMass spectrometry
■650 4▼aCloning
■650 4▼aDesign
■650 4▼aEngineering
■650 4▼aPhenols
■650 4▼aCell growth
■650 4▼aPolypeptides
■650 4▼aProteomics
■650 4▼aAnalytical chemistry
■650 4▼aBioinformatics
■650 4▼aCellular biology
■650 4▼aOrganic chemistry
■690 ▼a0389
■690 ▼a0459
■690 ▼a0537
■690 ▼a0486
■690 ▼a0715
■690 ▼a0379
■690 ▼a0490
■71020▼aStanford University.
■7730 ▼tDissertations Abstracts International▼g86-05B.
■790 ▼a0212
■791 ▼aPh.D.
■792 ▼a2024
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17164802▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.


