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Biosynthetic DNA-Protein Conjugation in Live Cells.
Biosynthetic DNA-Protein Conjugation in Live Cells.
상세정보
- 자료유형
- 학위논문
- Control Number
- 0017165068
- International Standard Book Number
- 9798346785286
- Dewey Decimal Classification Number
- 571.6
- Main Entry-Personal Name
- Verma, Shivam.
- Publication, Distribution, etc. (Imprint
- [S.l.] : Stanford University., 2024
- Publication, Distribution, etc. (Imprint
- Ann Arbor : ProQuest Dissertations & Theses, 2024
- Physical Description
- 103 p.
- General Note
- Source: Dissertations Abstracts International, Volume: 86-06, Section: B.
- General Note
- Advisor: Bertozzi, Carolyn.
- Dissertation Note
- Thesis (Ph.D.)--Stanford University, 2024.
- Summary, Etc.
- 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.
- Subject Added Entry-Topical Term
- Cell death.
- Subject Added Entry-Topical Term
- Acids.
- Subject Added Entry-Topical Term
- Antibodies.
- Subject Added Entry-Topical Term
- Mutation.
- Subject Added Entry-Topical Term
- Bacteria.
- Subject Added Entry-Topical Term
- Scientific imaging.
- Subject Added Entry-Topical Term
- E coli.
- Subject Added Entry-Topical Term
- Genotype & phenotype.
- Subject Added Entry-Topical Term
- Cell culture.
- Subject Added Entry-Topical Term
- Annealing.
- Subject Added Entry-Topical Term
- Protein synthesis.
- Subject Added Entry-Topical Term
- Plasmids.
- Subject Added Entry-Topical Term
- Mass spectrometry.
- Subject Added Entry-Topical Term
- Information storage.
- Subject Added Entry-Topical Term
- Empowerment.
- Subject Added Entry-Topical Term
- Microscopy.
- Subject Added Entry-Topical Term
- Genetic engineering.
- Subject Added Entry-Topical Term
- DNA polymerase.
- Subject Added Entry-Topical Term
- Toxins.
- Subject Added Entry-Topical Term
- Analytical chemistry.
- Subject Added Entry-Topical Term
- Biochemistry.
- Subject Added Entry-Topical Term
- Bioengineering.
- Subject Added Entry-Topical Term
- Cellular biology.
- Subject Added Entry-Topical Term
- Genetics.
- Added Entry-Corporate Name
- Stanford University.
- Host Item Entry
- Dissertations Abstracts International. 86-06B.
- Electronic Location and Access
- 로그인을 한후 보실 수 있는 자료입니다.
- Control Number
- joongbu:655758
MARC
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■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[S.l.]▼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.
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