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Robust ZFN-mediated genome editing in adult hemophilic mice.

ABSTRACT: Monogenic diseases, including hemophilia, represent ideal targets for genome-editing approaches aimed at correcting a defective gene. Here we report that systemic adeno-associated virus (AAV) vector delivery of zinc finger nucleases (ZFNs) and corrective donor template to the predominantly quiescent livers of adult mice enables production of high levels of human factor IX in a murine model of hemophilia B. Further, we show that off-target cleavage can be substantially reduced while maintaining robust editing by using obligate heterodimeric ZFNs engineered to minimize unwanted cleavage attributable to homodimerization of the ZFNs. These results broaden the therapeutic potential of AAV/ZFN-mediated genome editing in the liver and could expand this strategy to other nonreplicating cell types.

SUBMITTER: Anguela XM

PROVIDER: S-EPMC3821724 | biostudies-literature | 2013 Nov

REPOSITORIES: biostudies-literature

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Robust ZFN-mediated genome editing in adult hemophilic mice.

Anguela Xavier M XM Sharma Rajiv R Doyon Yannick Y Miller Jeffrey C JC Li Hojun H Haurigot Virginia V Rohde Michelle E ME Wong Sunnie Y SY Davidson Robert J RJ Zhou Shangzhen S Gregory Philip D PD Holmes Michael C MC High Katherine A KA

Blood 20131001 19

Monogenic diseases, including hemophilia, represent ideal targets for genome-editing approaches aimed at correcting a defective gene. Here we report that systemic adeno-associated virus (AAV) vector delivery of zinc finger nucleases (ZFNs) and corrective donor template to the predominantly quiescent livers of adult mice enables production of high levels of human factor IX in a murine model of hemophilia B. Further, we show that off-target cleavage can be substantially reduced while maintaining r ...[more]

PMID: 24085764

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