This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Adenoviruses are nonenveloped DNA viruses with icosahedral symmetry and ~900? in diameter. Their association with acute respiratory, gastrointestinal and ocular infections as well as use in gene delivery applications has drawn intense interest from basic and medical researchers. Approximately 28% of all viral vector gene therapy trials in the U.S. currently employ adenovirus, primarily for the treatment of cardiovascular diseases and cancer. Adenovirus also continues to serve as a useful tool for dissecting fundamental cell and molecular processes such as RNA slicing, cell cycle regulation and cancer. Currently we have identified several conditions that reproducibly allow the growth of 100-250 uM Ad crystals. Some of these crystals diffract to 8? at Biocars bealine BMC14 at APS. Any significant data collection on these large unit cell (>1000?) crystals can be done only at synchrotron sources, like APS, where focused X-ray beam with high flux and large 3x3 size CCD detectors are available. The helium cone and large size (3x3) detectors and capabilities to offset the detector are essential for collection of usable x-ray data to higher resolutions without the significant loss of reflection intensities. The knowledge gained from these structural studies should increase our understanding of nonenveloped virus assembly, cell entry and receptor interactions. They may also allow the development of modified Ad vectors for improved uses in gene therapy. Furthermore, we stronglybelieve that the successful determination of adenovirus structure at high resolution will be ahuge step forward in the pushing the ?size? limits using x-ray crystallography.
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