TY - JOUR TI - Targeted genome editing by lentiviral protein transduction of zinc-finger and TAL-effector nucleases AU - Cai, Yujia AU - Bak, Rasmus O AU - Mikkelsen, Jacob Giehm A2 - Golub, Todd VL - 3 PY - 2014 DA - 2014/04/24 SP - e01911 C1 - eLife 2014;3:e01911 DO - 10.7554/eLife.01911 UR - https://doi.org/10.7554/eLife.01911 AB - Future therapeutic use of engineered site-directed nucleases, like zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), relies on safe and effective means of delivering nucleases to cells. In this study, we adapt lentiviral vectors as carriers of designer nuclease proteins, providing efficient targeted gene disruption in vector-treated cell lines and primary cells. By co-packaging pairs of ZFN proteins with donor RNA in ‘all-in-one’ lentiviral particles, we co-deliver ZFN proteins and the donor template for homology-directed repair leading to targeted DNA insertion and gene correction. Comparative studies of ZFN activity in a predetermined target locus and a known nearby off-target locus demonstrate reduced off-target activity after ZFN protein transduction relative to conventional delivery approaches. Additionally, TALEN proteins are added to the repertoire of custom-designed nucleases that can be delivered by protein transduction. Altogether, our findings generate a new platform for genome engineering based on efficient and potentially safer delivery of programmable nucleases. KW - protein transduction KW - zinc-finger nuclease KW - transcription activator-like effector nuclease KW - lentiviral vector KW - Gag JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -