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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...
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  
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MARC

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■1001  ▼aCheah,  Joleen.
■24510▼aEngineering  Proximity  Labeling  Techniques  to  Map  Translocating  Proteomes  and  Rna  Reporters  to  Surveil  Cell  States
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■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.

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