Background: Three-dimensional (3D) visualization systems, also known as heads-up systems, are now available for eye surgery and as with every new device there is need for a specific evaluation. Objectives: The aim of this study was to compare the efficiency, surgical comfort, and safety of a 3D visualization system to a standard binocular microscope (BM) in routine ophthalmologic procedures. Method: After a 4-week training period, a 3D visualization system (Ngenuity, Alcon ®) available in one of the Robert Debré Hospital Ophthalmology Departments' operating rooms was compared to a standard BM (OPMI LUMIRA 700, Zeiss ®), in the process of a call for new device evaluation. From December 2017 to March 2018, 5 surgeons and their respective residents were asked to fill in a questionnaire for all procedures. Before the surgery, the surgeon recorded: (i) the type of surgery (cataract [PK], retinal detachment [RD], epiretinal membrane peeling [ERM], macular hole, vitreous haemorrhage [VH]), (ii) the type of visualization system chosen (3D or BM), and (iii) the estimated surgical risk (low, intermediate, or high grade). At the end of the procedure, the primary surgeon recorded
Gene targeting can be achieved with lentiviral vectors delivering donor sequences along with a nuclease that creates a locus-specific double-strand break (DSB). Therapeutic applications of this system would require an appropriate control of the amount of endonuclease delivered to the target cells, and potentially toxic sustained expression must be avoided. Here, we show that the nuclease can be transferred into cells as a protein associated with a lentiviral vector particle. I-SceI, a prototypic meganuclease from yeast, was incorporated into the virions as a fusion with Vpr, an HIV accessory protein. Integration-deficient lentiviral vectors containing the donor sequences and the I-SceI fusion protein were tested in reporter cells in which targeting events were scored by the repair of a puromycin resistance gene. Molecular analysis of the targeted locus indicated a 2-fold higher frequency of the expected recombination event when the nuclease was delivered as a protein rather than encoded by a separate vector. In both systems, a proportion of clones displayed multiple integrated copies of the donor sequences, either as tandems at the targeted locus or at unrelated loci. These integration patterns were dependent upon the mode of meganuclease delivery, suggesting distinct recombination processes.
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