Easily Used Kits to Evolve Reagents that Covalently Tag and Inactivate Proteins
易于使用的试剂盒可进化出共价标记和灭活蛋白质的试剂
基本信息
- 批准号:10626917
- 负责人:
- 金额:$ 31.39万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2021
- 资助国家:美国
- 起止时间:2021-09-01 至 2025-05-31
- 项目状态:未结题
- 来源:
- 关键词:2019-nCoVAccelerationAcylationAddressAmidesAmino Acid SequenceAntibodiesAreaBindingBinding ProteinsBioinformaticsBiologicalBiologyBiomedical ResearchBrain MappingCOVID-19 detectionCOVID-19 pandemicCatalytic AntibodiesCatalytic RNACell surfaceCellsChemicalsChemistryClinical Laboratory Information SystemsCoronavirusDNADNA biosynthesisDNA sequencingDataDevelopmentDiagnosticDiseaseDisease ManagementDoxorubicinEnzymesEstersEvolutionFeedbackFoundationsGPC3 geneImageIn VitroIndiaInformation SystemsKnowledgeLaboratoriesLanguageLicensingLysineMalignant neoplasm of liverMeasuresMedicalMedical ResearchMedicineModificationMolecular EvolutionNational Institute of General Medical SciencesOrganic ChemistryOutcomePeptidesPerformancePharmaceutical PreparationsPhysiologicalProcessProtein EngineeringProtein Sequence AnalysisProteinsReactionReaction TimeReagentReproducibilityResearch PersonnelResectedRestRibosomal RNARoleScienceShipsSiteSodium ChlorideSpecificitySurgeonSynthesis ChemistrySystemTechnologyTemperatureTestingThermodynamicsToxinTranslatingVisionWorkacyl groupamino groupbiological systemscancer cellcell killingcombinatorial chemistrydesigndrug discoverygene synthesisgenetic informationin vivoinnovationinventionmeetingsmolecular recognitionmultiplex diagnosticsnew technologynext generation sequencingprotein functionsuccesssynthetic biologytechnology platformtooltumorweb-based tool
项目摘要
Easily Used Kits to Evolve Reagents that Covalently Tag and Inactivate Proteins
Foundation for Applied Molecular Evolution
Steven Benner
ABSTRACT
Under PAR-19-253, the NIGMS seeks new technologies that create a positive feedback loop that drives
science forward by allowing new questions to be asked and new discoveries to be made, which in turn drives
the development of new technologies. The Benner group has, for 30 years, contributed to this NIGMS vision,
developing new technologies for NextGen DNA sequencing and NextGen DNA synthesis, bioinformatic and
evolutionary analyses, dynamic combinatorial chemistry for drug discovery, protein engineering, and new
platforms that make multiplexed diagnostics easy, platforms used today to manage the COVID pandemic.
Here, we offer the NIGMS another transformative tool, a platform to allow researchers to choose a
protein target and create a reagent (an AEGISZyme) that chemically transforms bound proteins. Such
reagents have been sought for 40 years with only limited success.
We will focus on one transformation: AEGISZymes that add an acyl group to an amino group on a lysine of the
bound target, where the acylation reagent is an ester. This acylation may inactivate the targeted protein, allow-
ing researchers to test hypotheses about the role of that protein in biology. It may carry a payload which, when
internalized with the target protein, carry drugs or stabilized AEGISZymes into a cell. It may fluorescently tag
the protein to help clinicians cut away fluorescing cancer cells selectively as they resect a tumor.
To achieve this transformative and innovative outcome, we will apply laboratory in vitro evolution (LIVE) to
artificially expanded genetic information systems (AEGIS). The platform will be delivered by meeting 3 Aims:
Aim 1. We will use AEGIS-LIVE to deliver AEGISZymes that acylate lysines in target proteins with pass/fail
reaction times of <10 sec-1. To test this, we will create these AEGISZymes that use a co-substrate carrying an
ester group for three targets. Rates of the selected AEGISZymes will be quantitated, specificity will be metricked
against similar targets with slightly different amino acid sequences, and modification sites will be found,
Aim 2. We will use AEGIS-LIVE to deliver AEGISZymes that acylate lysines on researcher-chosen targets with
turnover, with pass/fail turnovers of >1000 and kcat/KM of >105 M-1 sec-1. Turnover rates will be metricked
under physiological and laboratory conditions, and correlated to duplex stability from thermodynamic data.
Aim 3. We will use AEGIS-LIVE to deliver mirror AEGISZymes that are stable in biological systems, including
transport into cells. This will allow AEGISZymes to be used in biological media, to support nanotrain toxin
delivery, and to set the stage to use these molecules in vivo.
Aim 4. We will test the scope of the platform to address design parameters, such as how long random regions
should be, how good loops are as full protein surrogates, and how sequence space is searched.
Last, to lay to rest any view that AEGIS-LIVE is too "cumbersome", we will create distributable kits that allow
their recipients to make their own AEGISZymes. This is the ultimate in authentication and reproducibility.
易于使用的试剂盒进化试剂共价标记和灭活蛋白质
项目成果
期刊论文数量(4)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
DNA Structure Design Is Improved Using an Artificially Expanded Alphabet of Base Pairs Including Loop and Mismatch Thermodynamic Parameters.
使用人工扩展的碱基对字母表(包括环和错配热力学参数)改进 DNA 结构设计。
- DOI:10.1101/2023.06.06.543917
- 发表时间:2023
- 期刊:
- 影响因子:0
- 作者:Pham,TuanM;Miffin,Terrel;Sun,Hongying;Sharp,KennethK;Wang,Xiaoyu;Zhu,Mingyi;Hoshika,Shuichi;Peterson,RaymondJ;Benner,StevenA;Kahn,JasonD;Mathews,DavidH
- 通讯作者:Mathews,DavidH
Enzyme-Assisted High Throughput Sequencing of an Expanded Genetic Alphabet at Single Base Resolution.
单碱基分辨率的扩展遗传字母表的酶辅助高通量测序。
- DOI:10.21203/rs.3.rs-3678081/v1
- 发表时间:2023
- 期刊:
- 影响因子:0
- 作者:Wang,Bang;Bradley,KevinM;Kim,Myong-Jung;Laos,Roberto;Chen,Cen;Gerloff,DietlindL;Manfio,Luran;Yang,Zunyi;Benner,StevenA
- 通讯作者:Benner,StevenA
Functional Selection of Tau Oligomerization-Inhibiting Aptamers.
Tau 寡聚化抑制适体的功能选择。
- DOI:10.1002/anie.202402007
- 发表时间:2024
- 期刊:
- 影响因子:0
- 作者:Wang,Bang;Pan,Xiaoshu;Teng,I-Ting;Li,Xiaowei;Kobeissy,Firas;Wu,Zo-Yu;Zhu,Jiepei;Cai,Guangzheng;Yan,He;Yan,Xin;Liang,Mingwei;Yu,Fahong;Lu,Jianrong;Yang,Zunyi;Biondi,Elisa;Haskins,William;Cao,YCharles;Benner,StevenA;Tan
- 通讯作者:Tan
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STEVEN A BENNER其他文献
STEVEN A BENNER的其他文献
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{{ truncateString('STEVEN A BENNER', 18)}}的其他基金
Basic Research for Diagnostics and Surveillance in Lower Resource Environments
低资源环境诊断和监测基础研究
- 批准号:
10669039 - 财政年份:2021
- 资助金额:
$ 31.39万 - 项目类别:
Basic Research for Diagnostics and Surveillance in Lower Resource Environments
低资源环境诊断和监测基础研究
- 批准号:
10468606 - 财政年份:2021
- 资助金额:
$ 31.39万 - 项目类别:
Easily Used Kits to Evolve Reagents that Covalently Tag and Inactivate Proteins
易于使用的试剂盒可进化出共价标记和灭活蛋白质的试剂
- 批准号:
10478279 - 财政年份:2021
- 资助金额:
$ 31.39万 - 项目类别:
Easily Used Kits to Evolve Reagents that Covalently Tag and Inactivate Proteins
易于使用的试剂盒可进化出共价标记和灭活蛋白质的试剂
- 批准号:
10298982 - 财政年份:2021
- 资助金额:
$ 31.39万 - 项目类别:
Equipment Supplement to 1R01GM141391-01A1 (Easily Used Kits to Evolve Reagents that Covalently Tag and Inactivate Proteins)
1R01GM141391-01A1 的设备补充(易于使用的试剂盒,用于进化共价标记和灭活蛋白质的试剂)
- 批准号:
10580301 - 财政年份:2021
- 资助金额:
$ 31.39万 - 项目类别:
PHS2019-02 Omnibus Solic of the NIH, CDC, and FDA for SBIR Apps No Clinical Trial (Parent SBIR R43/4
PHS2019-02 NIH、CDC 和 FDA 的 SBIR 应用程序综合 Solic 尚未进行临床试验(母公司 SBIR R43/4
- 批准号:
10476977 - 财政年份:2021
- 资助金额:
$ 31.39万 - 项目类别:
Reagents to Chemically Tag Specific Coronavirus Spike Proteins
化学标记特定冠状病毒刺突蛋白的试剂
- 批准号:
10259048 - 财政年份:2021
- 资助金额:
$ 31.39万 - 项目类别:
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