Detecting DNA damage at a single-molecule level by atomic force microscopy
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چکیده
DNA damage can be caused daily by as many as one million molecular lesions per cell by normal metabolic activities and environmental factors. The uncorrected lesions in critical genes (such as tumor suppressor genes) can inhibit a cell's ability to carry out its regular function and accordingly increase the possibility of tumor formation. Detecting DNA damage is the first step towards understanding the complex damage repair process. Hereby, we introduce single-molecule methods to detect DNA damage using atomic force microscope (AFM) imaging and single-molecule force spectroscopy (SMFS). These new techniques allow us to visualize and manipulate single DNA under nearly in vivo conditions in real time. First, the majority of most DNA damage affects the primary structure of the double helix. The lesions cause a change in DNA mechanics which can be measured easily by SMFS. Second, a variety of repair strategies have evolved to restore lost genetic information. Specific DNA repair enzymes or antibodies bind at or near the site of damage, recruiting other molecules to bind and triggering the actual repair to take place. These processes can also be visualized directly, and the lesions can be quantified at a single-molecule level by AFM imaging of these repair enzymes or antibodies. The article highlights research progress in the detection of DNA damages using AFM imaging and SMFS methods. Multiple examples are presented on visualizing and manipulating DNA at a single-molecule level.
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تاریخ انتشار 2011