Cytotoxicity of cisplatin and mitomycin C (MMC) is ascribed largely to their ability to generate interstrand crosslinks (ICLs) in DNA, which block the progression of replication forks. The processing of ICLs requires the Fanconi anemia (FA) pathway, excision repair, and translesion DNA synthesis (TLS). It also requires homologous recombination (HR), which repairs double-strand breaks (DSBs) gen...

Biased integration remains a key challenge for gene therapy based on lentiviral vector technologies. Engineering of next-generation lentiviral vectors targeting safe genomic harbors for insertion is therefore of high relevance. In a previous paper (Cai et al., 2014a), we showed the use of integrase-defective lentiviral vectors (IDLVs) as carriers of complete gene repair kits consisting of zinc-...

The nuclease domain of ColE7 (N-ColE7) contains an H-N-H motif that folds in a beta beta alpha-metal topology. Here we report the crystal structures of a Zn2+-bound N-ColE7 (H545E mutant) in complex with a 12-bp duplex DNA and a Ni2+-bound N-ColE7 in complex with the inhibitor Im7 at a resolution of 2.5 A and 2.0 A, respectively. Metal-dependent cleavage assays showed that N-ColE7 cleaves doubl...

Genome editing technology revolutionized crop improvement through sequence-specific, precise, site-directed, safe genetic manipulation and combat the major 21st century challenge such as achieving world food security meeting rising global demand improving nutrition in face of rapidly changing climate conditions. Crop using conventional molecular breeding approaches takes time, causing biosafety...

Engineered nucleases, which incise the genome at predetermined sites, have a number of laboratory and clinical applications. There is, however, a need for better methods for controlled intracellular delivery of nucleases. Here, we demonstrate a method for ligand-mediated delivery of zinc finger nucleases (ZFN) proteins using transferrin receptor-mediated endocytosis. Uptake is rapid and efficie...

Zinc finger proteins acquire DNA-binding ability by Zn (II) complexation. In the zinc finger domain of the Cys2His2 type, each finger is approximately 30 amino acid residues long and consists of a simple ββα–fold stabilized by chelation of a zinc ion with the conserved Cys2His2 residues. A zinc finger motif of Cys2His2 offers an attractive framework for the design of a novel DNA-binding protein...