DNA helicases and associated factors in genome stability
DNA 解旋酶和基因组稳定性的相关因素
基本信息
- 批准号:10788644
- 负责人:
- 金额:$ 22.24万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2019
- 资助国家:美国
- 起止时间:2019-08-01 至 2024-07-31
- 项目状态:已结题
- 来源:
- 关键词:AllelesBiochemicalBiological AssayBiological ModelsBiophysicsChemistryChromatographyClinicCustomDNA Interstrand Cross-Link RepairDNA RepairDNA biosynthesisDataDedicationsDevelopmentDevicesDiseaseFamilyFunctional disorderFundingGenesGeneticGenetic RecombinationGenomeGenome StabilityGoalsHelicase GeneHumanIn VitroKnowledgeLinkMaintenanceMass Spectrum AnalysisMeasurementMolecular Sieve ChromatographyMutateMutationPatientsPhysiologicalPreparationProcessProteinsRecombinant ProteinsResearchResolutionRoleSystemTechniquesTelomere MaintenanceWorkcrosslinkdesignexperimental studygenome integrityhelicaseimprovedin vivoinsightmodel organismmutantnext generation sequencingrepairedsingle moleculetherapeutic targetvirtual
项目摘要
PROJECT SUMMARY
DNA helicases function in virtually all aspects of DNA replication, recombination, and repair. As such,
they are vital to maintaining genome integrity and are disease linked when mutated. Despite many in vivo and
in vitro advances in working with helicases, there is a gap in knowledge connecting mutant alleles of helicase
genes to the treatment of patients in clinics. The objective of my research is to gain mechanistic insight into
how DNA helicases function in genome maintenance and why their dysfunction leads to disease. Toward this
goal, we are studying PIF1 and RecQ family helicases, which are evolutionarily conserved and because
mutations in the human genes encoding these helicases are associated with multiple diseases. Our current
work focuses on the roles of RecQ helicases in DNA inter-strand crosslink (ICL) repair and RecQ and Pif1
helicases in telomere maintenance. To perform this work, we will employ a variety of classic and cutting edge
experimental techniques, from standard in vitro enzymatic assays and model organism genetics to next-
generation sequencing, crosslinking mass spectrometry, and the development of custom click chemistry
probes. Overall, this work will provide fundamental data critical to understanding how PIF1 and RecQ family
helicases aid in the maintenance of genome stability, and it will ultimately lead to therapeutic targets and
treatments for helicase-linked diseases.
I am requesting funds to purchase a Refeyn TwoMP mass photometer. This device enables rapid mass
measurements of small volumes of dilute biomolecules, with single-molecule precision. We routinely generate
recombinant protein preparations for biochemical assays, and knowing the oligomeric state and
monodispersity under physiological conditions is critical to the design, analysis, and interpretation of our
experiments. We currently use size-exclusion chromatography (SEC) to determine recombinant protein mass,
but SEC is slow, has low resolution, and requires large volumes of very concentrated protein. Therefore, SEC
limits the availability of our chromatography system and is incompatible with proteins that cannot be purified in
large amounts or do not tolerate the concentration process. The purchase of a mass photometer will provide a
rapid and high-resolution solution to these problems with a dedicated device that is inexpensive and easy to
use, improving our biophysical capabilities and supporting all of our projects.
项目摘要
DNA解旋酶几乎在DNA复制、重组和修复的所有方面发挥作用。因此,在本发明的一个方面,
它们对维持基因组完整性至关重要,突变时与疾病有关。尽管许多体内和
在体外研究解旋酶的进展中,连接解旋酶突变等位基因的知识存在空白
基因在临床上的应用我研究的目的是从机理上了解
DNA解旋酶如何在基因组维护中发挥作用以及为什么它们的功能障碍会导致疾病。朝着这个
我们正在研究PIF1和RecQ家族解旋酶,它们在进化上是保守的,
编码这些解旋酶的人类基因中的突变与多种疾病有关。我们目前
工作重点是RecQ解旋酶在DNA链间交联(ICL)修复中的作用,以及RecQ和Pif1
端粒维持中的解旋酶。为了完成这项工作,我们将聘请各种经典和尖端的
实验技术,从标准的体外酶测定和模式生物遗传学到下一个-
代测序、交联质谱和定制点击化学的发展
probes.总的来说,这项工作将提供关键的基本数据,以了解PIF1和RecQ家族
解旋酶有助于维持基因组的稳定性,并且它将最终导致治疗靶点,
解旋酶相关疾病的治疗。
我申请资金购买Refeyn TwoMP质量光度计。这个装置可以快速地
测量小体积的稀释生物分子,具有单分子精度。我们通常会生成
用于生物化学测定的重组蛋白制剂,并了解寡聚状态和
生理条件下的单分散性对于我们的设计、分析和解释是至关重要的。
实验我们目前使用尺寸排阻色谱法(SEC)来确定重组蛋白质质量,
但SEC速度慢,分辨率低,并且需要大量非常浓缩的蛋白质。因此,SEC
限制了我们的层析系统的可用性,并且与不能在层析系统中纯化的蛋白质不相容。
大量或不耐受浓缩过程。购买质量光度计将提供
快速和高分辨率的解决方案,这些问题的专用设备,是廉价的,易于
使用,提高我们的生物物理能力,并支持我们所有的项目。
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)
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{{ truncateString('Matthew Linne Bochman', 18)}}的其他基金
DNA helicases and associated factors in genome stability
DNA 解旋酶和基因组稳定性的相关因素
- 批准号:
10810234 - 财政年份:2023
- 资助金额:
$ 22.24万 - 项目类别:
DNA helicases and associated factors in genome stability
DNA 解旋酶和基因组稳定性的相关因素
- 批准号:
10388769 - 财政年份:2019
- 资助金额:
$ 22.24万 - 项目类别:
DNA helicases and associated factors in genome stability
DNA 解旋酶和基因组稳定性的相关因素
- 批准号:
10223373 - 财政年份:2019
- 资助金额:
$ 22.24万 - 项目类别:
DNA helicases and associated factors in genome stability
DNA 解旋酶和基因组稳定性的相关因素
- 批准号:
9796664 - 财政年份:2019
- 资助金额:
$ 22.24万 - 项目类别:
DNA helicases and associated factors in genome stability
DNA 解旋酶和基因组稳定性的相关因素
- 批准号:
10447062 - 财政年份:2019
- 资助金额:
$ 22.24万 - 项目类别:
DNA helicases and associated factors in genome stability
DNA 解旋酶和基因组稳定性的相关因素
- 批准号:
10680875 - 财政年份:2019
- 资助金额:
$ 22.24万 - 项目类别:
DNA helicases and associated factors in genome stability
DNA 解旋酶和基因组稳定性的相关因素
- 批准号:
10683104 - 财政年份:2019
- 资助金额:
$ 22.24万 - 项目类别:
DNA helicases and associated factors in genome stability
DNA 解旋酶和基因组稳定性的相关因素
- 批准号:
10644556 - 财政年份:2019
- 资助金额:
$ 22.24万 - 项目类别:
DNA helicases and associated factors in genome stability
DNA 解旋酶和基因组稳定性的相关因素
- 批准号:
10726902 - 财政年份:2019
- 资助金额:
$ 22.24万 - 项目类别:
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