Skip to main content
. 2009 Jan 19;37(5):1650–1662. doi: 10.1093/nar/gkp004

Figure 6.

Figure 6.

mCreI and mMsoI induce catalytic activity-dependent recombination in human cells. (a) In vivo recombination activity was assayed by co-transfecting coding plasmids for I-CreI or I-MsoI proteins together with a recombination reporter plasmid that contains a direct repeat of two genetically inactive copies of GFP. In vivo cleavage of the I-CreI or I-MsoI target site initiates gene conversion and repair of the cleaved copy to generate GFP+ cells that can be detected and quantified by flow cytometry (Materials and methods section). (b) Flow histograms of cells mock-transfected, transfected with reporter plasmid alone, or co-transfected with reporter: endonuclease plasmid pairs. The gate for GFP+ cells is shown by the boxed area, and the GFP+ frequency is given in the lower right of each histogram. (c) Frequency and fold increase in GFP+ cells for different reporter/coding plasmid combination. The frequency of GFP+ cells generated by in vitro XhoI-linearized DR-GFPCre reporter DNA (*) indicates that a substantial fraction of reporter molecules are likely cleaved in vivo by I-CreI and mCreI. D20N I-CreI and D22N I-MsoI are catalytically inactive mutants of I-CreI and I-MsoI which fail to induce GFP+ cells when cotransfected with a Cre or Mso-specific reporter plasmid. Error bars are means ± SDs.