| 摘要 |
The present invention provides compositions and methods concerning nucleic acid binding polypeptides, such as homing endonucleases, which are modified to possess sequencespecific activity at a modified target site. Methods for creating such modified polypeptides involves computer modeling either a polypeptide with a nucleic acid binding domain and a modified target nucleic acid sequence or a fusion of two nucleic acid binding domains (chimera) and a target nucleic acid sequence. Subsequent steps involve modification of the polypeptide or chimera to generate a protein that has site-specific activity at a particular target nucleic acid sequence. In some embodiments, the altered polypeptide has nuclease activity that is specific to the target site. In additional embodiments, the methods and compositions specifically concern homing endonucleases, such as the LAGLIDADG or His-Cys Box Group I homing endonucleases. |
