본문

서브메뉴

Biosynthetic DNA-Protein Conjugation in Live Cells
Biosynthetic DNA-Protein Conjugation in Live Cells
Biosynthetic DNA-Protein Conjugation in Live Cells

상세정보

자료유형  
 학위논문 서양
최종처리일시  
20250211153119
ISBN  
9798346785286
DDC  
571.6
저자명  
Verma, Shivam.
서명/저자  
Biosynthetic DNA-Protein Conjugation in Live Cells
발행사항  
[Sl] : Stanford University, 2024
발행사항  
Ann Arbor : ProQuest Dissertations & Theses, 2024
형태사항  
103 p
주기사항  
Source: Dissertations Abstracts International, Volume: 86-06, Section: B.
주기사항  
Advisor: Bertozzi, Carolyn.
학위논문주기  
Thesis (Ph.D.)--Stanford University, 2024.
초록/해제  
요약Conjugating DNA to protein has accelerated biotechnologies including high-throughput protein screening, super-resolution microscopy, and ultrasensitive diagnostics. Current approaches to synthesize DNA-protein conjugates (DPCs) are, however, limited in throughput. Here, we present a new method to generate DPCs entirely with biosynthetic machinery in live cells. We synthesize DPCs via fusion to bacterial HUH endonucleases, tyrosine autoconjugases that react with single-stranded DNA (ssDNA). The reactant ssDNA is produced in cells by repurposing bacterial retrons, specialized reverse transcriptases paired with template RNA. We start by reacting HUH endonucleases with retron ssDNA in live E. coli to produce biosynthetic DPCs for the first time. By discovering key factors degrading designer DPCs, we also study how native DPCs are processed endogenously. Next, we show that HUH endonucleases and retrons are active in the mammalian cell cytoplasm. Combining these pathways in human cells presents exciting opportunities to generate well-folded and post-translationally modified human DPCs. We believe that, in the near future, biosynthetic DPC synthesis could be developed into a high-throughput, pooled conjugation platform for diverse impact in future biotechnologies.
일반주제명  
Cell death
일반주제명  
Acids
일반주제명  
Antibodies
일반주제명  
Mutation
일반주제명  
Bacteria
일반주제명  
Scientific imaging
일반주제명  
E coli
일반주제명  
Genotype & phenotype
일반주제명  
Cell culture
일반주제명  
Annealing
일반주제명  
Protein synthesis
일반주제명  
Plasmids
일반주제명  
Mass spectrometry
일반주제명  
Information storage
일반주제명  
Empowerment
일반주제명  
Microscopy
일반주제명  
Genetic engineering
일반주제명  
DNA polymerase
일반주제명  
Toxins
일반주제명  
Analytical chemistry
일반주제명  
Biochemistry
일반주제명  
Bioengineering
일반주제명  
Cellular biology
일반주제명  
Genetics
기타저자  
Stanford University.
기본자료저록  
Dissertations Abstracts International. 86-06B.
전자적 위치 및 접속  
로그인 후 원문을 볼 수 있습니다.

MARC

 008250123s2024        us                              c    eng  d
■001000017165068
■00520250211153119
■006m          o    d                
■007cr#unu||||||||
■020    ▼a9798346785286
■035    ▼a(MiAaPQ)AAI31757691
■035    ▼a(MiAaPQ)Stanfordyx012pw2225
■040    ▼aMiAaPQ▼cMiAaPQ
■0820  ▼a571.6
■1001  ▼aVerma,  Shivam.
■24510▼aBiosynthetic  DNA-Protein  Conjugation  in  Live  Cells
■260    ▼a[Sl]▼bStanford  University▼c2024
■260  1▼aAnn  Arbor▼bProQuest  Dissertations  &  Theses▼c2024
■300    ▼a103  p
■500    ▼aSource:  Dissertations  Abstracts  International,  Volume:  86-06,  Section:  B.
■500    ▼aAdvisor:  Bertozzi,  Carolyn.
■5021  ▼aThesis  (Ph.D.)--Stanford  University,  2024.
■520    ▼aConjugating  DNA  to  protein  has  accelerated  biotechnologies  including  high-throughput  protein  screening,  super-resolution  microscopy,  and  ultrasensitive  diagnostics.  Current  approaches  to  synthesize  DNA-protein  conjugates  (DPCs)  are,  however,  limited  in  throughput.  Here,  we  present  a  new  method  to  generate  DPCs  entirely  with  biosynthetic  machinery  in  live  cells.  We  synthesize  DPCs  via  fusion  to  bacterial  HUH  endonucleases,  tyrosine  autoconjugases  that  react  with  single-stranded  DNA  (ssDNA).  The  reactant  ssDNA  is  produced  in  cells  by  repurposing  bacterial  retrons,  specialized  reverse  transcriptases  paired  with  template  RNA.  We  start  by  reacting  HUH  endonucleases  with  retron  ssDNA  in  live  E.  coli  to  produce  biosynthetic  DPCs  for  the  first  time.  By  discovering  key  factors  degrading  designer  DPCs,  we  also  study  how  native  DPCs  are  processed  endogenously.  Next,  we  show  that  HUH  endonucleases  and  retrons  are  active  in  the  mammalian  cell  cytoplasm.  Combining  these  pathways  in  human  cells  presents  exciting  opportunities  to  generate  well-folded  and  post-translationally  modified  human  DPCs.  We  believe  that,  in  the  near  future,  biosynthetic  DPC  synthesis  could  be  developed  into  a  high-throughput,  pooled  conjugation  platform  for  diverse  impact  in  future  biotechnologies.
■590    ▼aSchool  code:  0212.
■650  4▼aCell  death
■650  4▼aAcids
■650  4▼aAntibodies
■650  4▼aMutation
■650  4▼aBacteria
■650  4▼aScientific  imaging
■650  4▼aE  coli
■650  4▼aGenotype  &  phenotype
■650  4▼aCell  culture
■650  4▼aAnnealing
■650  4▼aProtein  synthesis
■650  4▼aPlasmids
■650  4▼aMass  spectrometry
■650  4▼aInformation  storage
■650  4▼aEmpowerment
■650  4▼aMicroscopy
■650  4▼aGenetic  engineering
■650  4▼aDNA  polymerase
■650  4▼aToxins
■650  4▼aAnalytical  chemistry
■650  4▼aBiochemistry
■650  4▼aBioengineering
■650  4▼aCellular  biology
■650  4▼aGenetics
■690    ▼a0486
■690    ▼a0487
■690    ▼a0202
■690    ▼a0379
■690    ▼a0369
■71020▼aStanford  University.
■7730  ▼tDissertations  Abstracts  International▼g86-06B.
■790    ▼a0212
■791    ▼aPh.D.
■792    ▼a2024
■793    ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17165068▼nKERIS▼z이  자료의  원문은  한국교육학술정보원에서  제공합니다.

미리보기

내보내기

chatGPT토론

Ai 추천 관련 도서


    신착도서 더보기
    최근 3년간 통계입니다.

    소장정보

    • 예약
    • 소재불명신고
    • 나의폴더
    • 우선정리요청
    • 비도서대출신청
    • 야간 도서대출신청
    소장자료
    등록번호 청구기호 소장처 대출가능여부 대출정보
    TF12670 전자도서 대출가능 마이폴더 부재도서신고 비도서대출신청 야간 도서대출신청

    * 대출중인 자료에 한하여 예약이 가능합니다. 예약을 원하시면 예약버튼을 클릭하십시오.

    해당 도서를 다른 이용자가 함께 대출한 도서

    관련 인기도서

    로그인 후 이용 가능합니다.