Fig. 5

ZFN and heat-inducible I-SceI activity. a Sequences of CCR5 site amplified by PCR using e – f primers (see Fig. 4a). Sequences obtained from the parental S1 line and four UZ lines (transformed with pUbi:ZFN). Deletion is shown by dots and insertion by small case letters. Red fonts indicate CCR5 sequence; b PCR analysis of UZ lines with g – f primers to detect large deletions at CCR5 site. Note the presence of <2.1 kb bands in different lanes; c PCR analysis of HI lines (1–3, S1 line retransformed with heat-inducible I-SceI gene). HI samples maintained at room temperature (RT) or exposed to heat-shock (HS) were analyzed to detect the excision of Bar gene using primers a2 - b (see Fig. 3a). Note the presence of 1.8 kb excision footprint in HS samples; d Sequences of 1.8 kb excision footprints from HS samples. Red fonts indicate I-SceI target sequences