权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

GenSyn: A Cycle for Serial Assembly of Very Large DNA Molecules

GenSyn：非常大的 DNA 分子的串行组装循环

基本信息

批准号：
10552680
负责人：
DAVID C. SCHWARTZ
金额：
$ 26.55万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
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：
2022-01-19 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10552680
关键词：
Adsorption Attenuated Bacterial Artificial Chromosomes Binding Biological Assay Buffers Cells Charge Chemicals Chromosomes Complement Computer Models Computer Simulation Coupling DNA DNA Binding DNA Ligation DNA biosynthesis Derivation procedure Device Designs Dimensions Engineering Fluorescence Genome Genomic DNA Genomics Glass Grain Health Human Human Genome Image Image Analysis In Vitro Ionic Strengths Laboratories Length Ligation Maps Measurement Mediating Methods Microscopy Modality Modeling Modification Molecular Molecular Conformation Names Nucleic Acids Oligonucleotides Optics Pathway interactions Peptides Performance Periodicity Phase Physical condensation Polyamines Process Reagent Reproducibility Research Scheme Solid Stretching Surface System Techniques Technology Validation copolymer design electric field fabrication foot manufacture microscopic imaging molecular modeling monomer novel operation physical property polyacrylamide polyacrylamide hydrogels research and development response single molecule synergism synthetic construct synthetic nucleic acid technology development

项目摘要

Abstract The broad, long-term objectives of the proposed research are development of a system for direct, cell-free, validated fabrication of very large DNA assemblies, for applications in synthetic applications and genome assembly. This system – “GenSyn” – will harness the advantages of large DNAs for synthesis on supports in ways that will complement and synergize cell-based techniques for the serial assembly of very large DNA molecules. The proposed research is based on i) binding approaches supporting chemical and enzymatic processes; ii) shear-free fluidic operations; iii) new methods for enhancing hybridization/ligation yields and iv) validations. The devices are designed for gentle fluidic operations supporting assembly and image analysis of large DNA constructs by epifluorescence microscopy. The first step in the GenSyn cycle will be binding DNA molecules to a support by hybridization with oligonucleotides that are bound to a derivatized glass surface. The hybridization and ligation steps will be optimized using physical and chemical manipulations. Modified large DNAs will be used as coupler molecules, supporting GenSyn cycles. Overall cycle performance metrics (coupling yields as a function of cycle number, molecule breakage, reagent retention, timings and enzymatic activity) will be assessed in ways that will inform and be guided by computer models. This research will help advance fundamental studies on genomes and human health.

摘要