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Peyronie’s disease can be extremely painful and even affect your
sleep. A “disease associated gene” is one that is defective in some
manner in a monogenic disease. In the methods of the
disclosure, one or more targeted nucleases as described herein create
a double-stranded break in the target sequence (e.g.,
cellular chromatin) at a predetermined site, and a “donor” polynucleotide, having homology to the nucleotide sequence in the region of the break,
can be introduced into the cell. Furthermore, the methods of targeted integration as described herein can also be used to integrate one
or more exogenous sequences. In certain embodiments, fusion polypeptides are used
for targeted double-stranded DNA cleavage. In other embodiments, the donor sequence is non-homologous to the first
sequence, and is inserted into the genome by non-homologous recombination mechanisms.
Thus, a first sequence in cellular chromatin can be altered and,
in certain embodiments, can be converted into a sequence
present in a donor polynucleotide. In any of the methods described herein, the first nucleotide
sequence (the “donor sequence”) can contain sequences that are homologous, but not identical, to genomic sequences in the region of interest, thereby stimulating homologous recombination to insert a non-identical sequence in the region of interest.
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