Nucleoprotein machines carry out essential biological processes including synthesis, modification, and repairof DNA and RNA. We propose to establish a nanomedicine development center (NDC) focusing on a modelnanomachine that carries out nonhomologous end joining (NHEJ) of DNA double strand breaks. This andother DNA repair machines have relatively simple structures (< 20 components) and significant biologicaland clinical relevance. DNA damage repair is vitally important to human health, as both normal metabolicactivities and environmental factors can cause DNA damage, resulting in as many as 100,000 individualmolecular lesions per cell per day. If allowed to accumulate without repair, these lesions interfere with genetranscription and replication, leading to premature aging, apoptosis, or unregulated cell division. We haveassembled an interdisciplinary team from eight institutions, with significant expertise in cell and molecularbiology of DNA damage repair, protein tagging and targeting, nanostructured probes, cryo-electronmicroscopy, signal-cell imaging, quantitative image analysis and computational biology, and light microscopyinstrumentation. We will develop innovative nanotechnologies and biomolecular approaches to elucidate thestructure-function relationships within and among DNA repair nanomachines. General principles emergingfrom these studies will lay a foundation for precise modification of the information stored in DNA and RNA,leading ultimately to novel therapeutic strategies for a wide range of diseases, including cancer. The NDChas five closely related aims including: (1) to develop orthogonal protein tagging strategies and novelfluorescence probes including quantum dot bioconjugates for nanomachine targeting; (2) to decipherstructure-function relationship of components required for the core NHEJ reaction; (3) to characterize thedynamics of nanomachine assembly and disassembly in the context of repair foci; (4) to determine thedimensions and structure of repair foci at high resolution in fixed cells; (5) to establish the engineering designprinciples underlying DNA double-strand break repair. This NDC will complement existing NDCs that focuson filaments, membranes and protein folding enzymes, and the probes, tools and methodologies developedwill be applicable to a wide range of biological and disease studies. Our long-term vision is to provide geneticcures for common human diseases based on the ability to manipulate the somatic human genome usingnanomedicine approaches that are inexpensive, effective, and user-friendly, similar to vaccination today.
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