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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
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■020 ▼a9798346785286
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■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.


