Methods of Generating Zinc Finger Nucleases Having Altered Activity
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United States of America Patent
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Issued Date -
Dec 27, 2012
app pub date -
Jan 21, 2011
filing date -
Jan 22, 2010
priority date (Note) -
Abandoned
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Abstract
Provided herein are zinc linger nucleases having altered, arid in particular, improved catalytic activity and methods of generating such nucleases. Accordingly, there are provided methods for identifying improved catalytic activity of a ZFN by expressing a mutated zinc finger nuclease in a cell containing a reporter construct with a toxic gene, and a zinc finger nuclease cleavage site that is recognized by the ZFN. Survival of the cell is positively correlated with catalytic activity of the ZFN; thus, libraries of mutated ZFKs may be selected for altered catalytic activity based on relative survival rates, Methods of using identified ZFNs are also provided.
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