Advanced search×

Analysis of illegitimate genomic integration mediated by zinc-finger nucleases: implications for specificity of targeted gene correction

Petter A Olsen, Monika Gelazauskaite, Markus Randøl, and Stefan Krauss

BMC Mol Biol 11(1):35 (2010) PMID 20459736

Formation of site specific genomic double strand breaks (DSBs), induced by the expression of a pair of engineered zinc-finger nucleases (ZFNs), dramatically increases the rates of homologous recombination (HR) between a specific genomic target and a donor plasmid. However, for the safe use of ZFN induced HR in practical applications, possible adverse effects of the technology such as cytotoxicity and genotoxicity need to be well understood. In this work, off-target activity of a pair of ZFNs has been examined by measuring the ratio between HR and illegitimate genomic integration in cells that are growing exponentially, and in cells that have been arrested in the G2/M phase.

DOI: 10.1186/1471-2199-11-35
Version: za2963e q8za2 q8zbb q8zc0 q8zd3 q8ze7 q8zf8 q8zg1
View PDF

Similar articles you may find interesting…

  1. Nontransgenic genome modification in plant cells.
    Ira I Marton, Amir A Zuker, ... Alexander A Vainstein

    Annu Rev Plant Physiol 154(3):1079-87 (2010) PMID 20876340

    We present an alternative, non-transgenic approach for ZFN delivery and production of mutant plants using a novel Tobacco rattle virus (TRV)-based expression system for indirect transient delivery of ZFNs into a variety of tissues and cells of intact plants. TRV systemically infected its hosts and v...
  2. Targeted integration in rat and mouse embryos with zinc-finger nucleases.
    Xiaoxia X Cui, Diana D Ji, ... Edward J EJ Weinstein

    Audio, Transactions of the IRE Professional... 29(1):64-7 (2011) PMID 21151125

    We use ZFN technology in embryos to introduce sequence-specific modifications (knock-ins) by means of homologous recombination in Sprague Dawley and Long-Evans hooded rats and FVB mice. This approach enables precise genome engineering to generate modifications such as point mutations, accurate inser...
  3. Correction for Osakabe et al., Site-directed mutagenesis in Arabidopsis using custom-designed zinc finger nucleases
    Author(s) unavailable

    Proc Natl Acad Sci U S A 108(1):433-433 (2011) PMID 21156829