Figure 5.

Genetic control of the survivor frequency and spectrum. (A) The two types of recurrent large deletions detected among surviving colonies. The 1.2 kb deletion as endpoints in 13-bp direct repeats (CCAAGCTACTACA), one of which overlaps the DNA binding site of the RyB subunit of the ZFN. The 11.7 kb deletion has endpoints in 14-bp direct repeats (TGGAAAAAAAAAAA) and removes the LYS2-flanking genes TKL2 and RAD16 (not shown). (B) Mean frequencies of surviving colonies that lost the ZFN cleavage site; error bars are 95% confidence intervals for the means. (C) The distributions of the three major survivor types are shown. Each mutant distribution was compared to the WT using a 2 × 3 contingency chi square test (ns, not significant). If the global p value (above each spectrum) was less than 0.05, individual mutation classes were compared using the Bonferroni correction to determine significance (p<0.05/3); asterisks indicate a significant proportional increase/decrease.