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Structural and Mechanistic Studies of CRISPR-Associated Transposons.
Structural and Mechanistic Studies of CRISPR-Associated Transposons.
상세정보
- 자료유형
- 학위논문
- Control Number
- 0017160223
- International Standard Book Number
- 9798384048275
- Dewey Decimal Classification Number
- 574
- Main Entry-Personal Name
- Tsai, Amy Wei-Lun.
- Publication, Distribution, etc. (Imprint
- [S.l.] : Cornell University., 2024
- Publication, Distribution, etc. (Imprint
- Ann Arbor : ProQuest Dissertations & Theses, 2024
- Physical Description
- 210 p.
- General Note
- Source: Dissertations Abstracts International, Volume: 86-03, Section: B.
- General Note
- Advisor: Kellogg, Elizabeth.
- Dissertation Note
- Thesis (Ph.D.)--Cornell University, 2024.
- Summary, Etc.
- 요약Current gene-editing tools, including CRISPR and transposon systems, serve various purposes for treating genetic diseases. However, they lack precision in inserting large DNA sequences into specific areas of interest. A novel class of genetic elements, CRISPR-associated transposons (CAST), were recently discovered to be capable of performing DNA insertions via RNA-guided transposition. CASTs merge the precise targeting capability of CRISPR systems with the ability to transport large DNA cargos from transposons, showing significant potential as an innovative gene editing tool.To realize the potential of these novel CAST systems, little is known regarding how elements of CAST systems are assembled and how those elements act together to perform DNA integration. This thesis describes efforts made to structurally characterize each protein component from a type V-K CAST from Scytonema Hofmanni (ShCAST) and to elucidate the underlying molecular mechanism through a combination of biochemical studies and single-particle cryogenic-electron microscopy (cryo-EM). Importantly, the structure of the transpososome, an integration complex containing all CAST components, is presented here. Insights provided by the structure of this complex are used to inform a comprehensive mechanistic model for the activity of the complex, including targeting and regulation. The model reveals how CAST proteins collectively act as a molecular machine to insert DNA at a genomic target site. The results from this study provide novel avenues for the rational optimization of CASTs for highly efficient gene insertions with single base-pair resolution, a critical goal in the field of gene therapy.
- Subject Added Entry-Topical Term
- Biology.
- Subject Added Entry-Topical Term
- Molecular biology.
- Subject Added Entry-Topical Term
- Genetics.
- Index Term-Uncontrolled
- CRISPR
- Index Term-Uncontrolled
- CRISPR-associated transposons
- Index Term-Uncontrolled
- Cryo-EM
- Index Term-Uncontrolled
- Structural biology
- Index Term-Uncontrolled
- Transposons
- Added Entry-Corporate Name
- Cornell University Chemistry and Chemical Biology
- Host Item Entry
- Dissertations Abstracts International. 86-03B.
- Electronic Location and Access
- 로그인을 한후 보실 수 있는 자료입니다.
- Control Number
- joongbu:654084
MARC
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■035 ▼a(MiAaPQ)AAI30991144
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a574
■1001 ▼aTsai, Amy Wei-Lun.▼0(orcid)0000-0001-8856-617X
■24510▼aStructural and Mechanistic Studies of CRISPR-Associated Transposons.
■260 ▼a[S.l.]▼bCornell University. ▼c2024
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2024
■300 ▼a210 p.
■500 ▼aSource: Dissertations Abstracts International, Volume: 86-03, Section: B.
■500 ▼aAdvisor: Kellogg, Elizabeth.
■5021 ▼aThesis (Ph.D.)--Cornell University, 2024.
■520 ▼aCurrent gene-editing tools, including CRISPR and transposon systems, serve various purposes for treating genetic diseases. However, they lack precision in inserting large DNA sequences into specific areas of interest. A novel class of genetic elements, CRISPR-associated transposons (CAST), were recently discovered to be capable of performing DNA insertions via RNA-guided transposition. CASTs merge the precise targeting capability of CRISPR systems with the ability to transport large DNA cargos from transposons, showing significant potential as an innovative gene editing tool.To realize the potential of these novel CAST systems, little is known regarding how elements of CAST systems are assembled and how those elements act together to perform DNA integration. This thesis describes efforts made to structurally characterize each protein component from a type V-K CAST from Scytonema Hofmanni (ShCAST) and to elucidate the underlying molecular mechanism through a combination of biochemical studies and single-particle cryogenic-electron microscopy (cryo-EM). Importantly, the structure of the transpososome, an integration complex containing all CAST components, is presented here. Insights provided by the structure of this complex are used to inform a comprehensive mechanistic model for the activity of the complex, including targeting and regulation. The model reveals how CAST proteins collectively act as a molecular machine to insert DNA at a genomic target site. The results from this study provide novel avenues for the rational optimization of CASTs for highly efficient gene insertions with single base-pair resolution, a critical goal in the field of gene therapy.
■590 ▼aSchool code: 0058.
■650 4▼aBiology.
■650 4▼aMolecular biology.
■650 4▼aGenetics.
■653 ▼aCRISPR
■653 ▼aCRISPR-associated transposons
■653 ▼aCryo-EM
■653 ▼aStructural biology
■653 ▼aTransposons
■690 ▼a0306
■690 ▼a0369
■690 ▼a0307
■71020▼aCornell University▼bChemistry and Chemical Biology.
■7730 ▼tDissertations Abstracts International▼g86-03B.
■790 ▼a0058
■791 ▼aPh.D.
■792 ▼a2024
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17160223▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
