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499 and 496 (numbered relative to wild-type FokI),
for instance mutations that replace the wild type Gln (Q) residue at position 486 with a Glu (E) residue, the wild type Iso (I)
residue at position 499 with a Leu (L) residue and the wild-type Asn (N) residue at position 496 with an Asp (D) or Glu (E) residue (also referred to as a "ELD" and "ELE" domains, respectively).
Additional engineered cleavage half-domains of FokI
that form obligate heterodimers can also be used in the ZFNs
described herein. In other embodiments, the nuclease comprises an engineered
TALE DNA-binding domain and a nuclease domain (e.g., endonuclease and/or meganuclease domain), also referred to as TALENs.
In other embodiments, the DNA-binding domain comprises a naturally occurring or engineered
(non-naturally occurring) TAL effector DNA binding domain. Thus, in some embodiments, the
DNA binding domain that binds to a target site in a target locus is an engineered
domain from a TAL effector similar to those derived from the plant pathogens Xanthomonas (see Boch et al,
(2009) Science 326: 1509-1512 and Moscou and Bogdanove, (2009)
Science 326: 1501) and Ralstonia (see Heuer et al (2007) Applied and Environmental Microbiology
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