Structural Basis of Programmable DNA-Insertion via Cryo-EM Studies of CRISPR-Associated TnsC
通过冷冻电镜研究 CRISPR 相关 TnsC 的可编程 DNA 插入的结构基础
基本信息
- 批准号:10543118
- 负责人:
- 金额:$ 32.86万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2022
- 资助国家:美国
- 起止时间:2022-01-01 至 2023-08-10
- 项目状态:已结题
- 来源:
- 关键词:ATP HydrolysisATP phosphohydrolaseAdoptionAreaBase PairingBehaviorBindingBinding SitesBiological ModelsCRISPR/Cas technologyClustered Regularly Interspaced Short Palindromic RepeatsCommunitiesComplementComplexCryoelectron MicroscopyDNADNA BindingDNA IntegrationDNA StructureDataElementsEngineeringEnvironmentEventExclusionExhibitsFilamentGenesGenome engineeringGoalsGuide RNAHealthHumanHydrolysisImmunityLinkLiteratureModelingMolecularMolecular ConformationNucleotidesOutcomeProcessProteinsRNA BindingRecruitment ActivityResolutionRoleSiteStructureSystemTransposaseds-DNAgenome editinginsightmolecular assembly/self assemblymu transposaseparticlepreferencereconstructionrecruittool
项目摘要
Project Summary
Recently, new macromolecular systems have been discovered which marries the benefits of both CRISPR
and TNP systems and shows tremendous promise as programmable DNA-insertion tools for genome-editing,
complementing the power of tools such as CRISPR-Cas9. This proposal aims to uncover the molecular
mechanisms governing two as-yet poorly understood phenomena in CRISPR-Transposase (CRISPR-TNP)
systems: target-site immunity and programmed-DNA insertion. The central protein thought to be responsible for
both of these observed behaviors in the multi-component shCAST system is shTnsC. We propose to utilize high-
resolution cryo-EM to determine the structure of DNA-bound shTnsC. In addition, shTnsC is a AAA+ ATPase
whose nucleotide-hydrolysis activity is linked to transposition. We propose to determine the structure of shTnsC
in different nucleotide-bound states in order to reveal the role of ATP-hydrolysis in transposition. Finally, we aim
to uncover the mechanisms governing shTnsC recruitment to the target-site via Cas12k and shTniQ, and how
the association between these factors ultimately initiates shCAST transposition. Our strong preliminary data
indicates that these aims are likely to be successful. In addition, the Kellogg lab is well-supported within the
Cornell community to achieve the goals outlined in this proposal.
项目摘要
最近,已经发现了新的大分子系统,其结合了CRISPR和CRISPR的优点。
和TNP系统,并显示出作为基因组编辑的可编程DNA插入工具的巨大前景,
补充了CRISPR-Cas9等工具的功能。这项提案旨在揭示
CRISPR转座酶(CRISPR-TNP)中两种尚未被充分理解的现象的机制
系统:靶点免疫和程序化DNA插入。被认为是负责
在多组分shCAST系统中观察到的这两种行为都是shTnsC。我们建议利用高-
分辨率cryo-EM以确定DNA结合的shTnsC的结构。此外,shTnsC是AAA+ ATP酶
其核苷酸水解活性与转座有关。我们建议确定shTnsC的结构
在不同的核苷酸结合状态,以揭示在转座ATP水解的作用。最后,我们的目标
揭示通过Cas 12 k和shTniQ将shTnsC募集到靶位点的机制,以及如何
这些因子之间的关联最终启动shCAST转座。我们强大的初步数据
这表明这些目标很可能会成功。此外,凯洛格实验室得到了
康奈尔社区实现本提案中概述的目标。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Elizabeth Kellogg其他文献
Elizabeth Kellogg的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Elizabeth Kellogg', 18)}}的其他基金
Structural Basis of Programmable DNA-Insertion via Cryo-EM Studies of CRISPR-Associated TnsC
通过冷冻电镜研究 CRISPR 相关 TnsC 的可编程 DNA 插入的结构基础
- 批准号:
10344519 - 财政年份:2022
- 资助金额:
$ 32.86万 - 项目类别:
Structural Basis of Programmable DNA-Insertion via Cryo-EM Studies of CRISPR-Associated TnsC
通过冷冻电镜研究 CRISPR 相关 TnsC 的可编程 DNA 插入的结构基础
- 批准号:
10797749 - 财政年份:2022
- 资助金额:
$ 32.86万 - 项目类别:
Cryo Transmission Electron Microscope for Cryo-EM Sample Optimization
用于冷冻电镜样品优化的冷冻透射电子显微镜
- 批准号:
10177173 - 财政年份:2021
- 资助金额:
$ 32.86万 - 项目类别:
Molecular Basis of Genome Organization and Integrity Using Cryo-EM
使用冷冻电镜研究基因组组织和完整性的分子基础
- 批准号:
10079493 - 财政年份:2017
- 资助金额:
$ 32.86万 - 项目类别:
Molecular Basis of Genome Organization and Integrity Using Cryo-EM
使用冷冻电镜研究基因组组织和完整性的分子基础
- 批准号:
9922323 - 财政年份:2017
- 资助金额:
$ 32.86万 - 项目类别:
Towards an understanding of telomere end protection: Cryo-EM studies of shelterin structure and function
了解端粒末端保护:Shelterin 结构和功能的冷冻电镜研究
- 批准号:
9371709 - 财政年份:2017
- 资助金额:
$ 32.86万 - 项目类别: