Fig. 1 | Nature Communications

Fig. 1

From: Diversifying the structure of zinc finger nucleases for high-precision genome editing

Fig. 1

Linkers and architectures developed in this study. a Sketch of the canonical ZFN dimer architecture. Circles marked with a scissors symbol denote the FokI cleavage domain. A tandem array of six arrows indicates each designed six-finger ZFP. Key features of this architecture include attachment of the FokI nuclease domain to the carboxy terminus of each zinc finger array and a lack of base-skipping between adjacent zinc fingers. ZFNs are shown interacting with duplex DNA, with black text on a gray background denoting ZFN target sites. b Alternative architectures enabled via pairing of ZFNs bearing an amino-terminal FokI cleavage domain (dark blue) and a carboxy-terminal FokI cleavage domain (light blue). The linker joining the FokI nuclease domain to the amino terminus of the ZFP is shown in red. ZFNs bearing an amino-terminal FokI attachment are able to recognize a target on the opposite DNA strand, relative to their canonical counterparts (compare with a). Thus, these architectures allow both ZFNs to recognize the same DNA strand. These two architectures are structurally identical, although for this study they will be referred to as NC and CN dimers denoting the FokI attachment point for the upstream and downstream ZFN, respectively. c ZFN architecture enabled via pairing two ZFNs with amino-terminal FokI nuclease domain fusions. This architecture is the inverse of the canonical pair shown in a and is referred to as an NN dimer. d Recognition of alternative DNA frames and sequences enabled by insertion of base-skipping linkers between fingers 2 and 3 or 4 and 5 of a six-finger ZFP. Skipped bases are shown without a gray background. The skipping linker is shown as a red bar between fingers

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