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. 2004 Nov;24(21):9305–9316. doi: 10.1128/MCB.24.21.9305-9316.2004

FIG. 1.

FIG. 1.

Reporters for measuring homologous DSB repair by SSA and HDR. (a) hprtSAGFP reporter. The structure of the reporter is shown before and after I-SceI cleavage and SSA. Note that repair by SSA results in a 2.7-kb deletion in the chromosome. Southern blot analysis is shown from an untransfected cell line with the SA-GFP reporter and from the same line after I-SceI expression and flow sorting to enrich for a pool of GFP+ cells. Large black triangles depict the 3′ end of the AFP cassette. (b) hprtDRGFP reporter. The structure of the reporter is shown before and after I-SceI cleavage and HDR. Note that repair by HDR converts the I-SceI site but otherwise maintains the structure of the reporter, as confirmed previously by Southern blot analysis (37). (c) Flow cytometric analysis of wild-type ES cells containing the hprtDRGFP and hprtSAGFP reporters targeted to the hprt locus, either untransfected or transfected with the I-SceI expression plasmid. Green fluorescence (FL1) is plotted on the y axis, with orange fluorescence (FL2) on the x axis.

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