It never fails—you want your best scissors, your roll of duct tape, and your finest pen, but they’re all in different rooms. The best tools do you no good at all if they’re not together, whether it’s in your house or in a plant cell. Recent work has generatedmany novel tools for genome engineering (reviewed in Belhaj et al., 2013). For example, engineered nucleases can generate site-specific do...

Site-specific and adaptable DNA binding domains are essential modules to develop genome engineering technologies for crop improvement. Transcription activator-like effectors (TALEs) proteins are used to provide a highly specific and adaptable DNA binding modules. TALE chimeric nucleases (TALENs) were used to generate site-specific double strand breaks (DSBs) in vitro and in yeast, Caenorhabditi...

Here, we examined the effects of molecular crowding on the function, structure and stability of nucleases. We found that the hydrolysis of a 29-mer double-stranded DNA by the endonucleases DNase I and S1 nuclease was substantially enhanced by molecular crowding using polyethylene glycol (PEG); however, molecular crowding had little effect on hydrolysis by exo III and exo I exonucleases. Moreove...