Mutational specificity: Mutation frequencies butnot mutant frequencies in Big Blue® mice fit a Poisson distribution

Transgenic mutation assays generally use mutant frequencies to estimate mutation frequencies but the degree to which clonal expansion inflates mutant frequencies is largely unknown. Mutant frequency is defined as the fraction of cells carrying mutations in the gene of interest and, according to the standard Big Blue protocol, is determined by dividing the number of mutant plaques by the total number of plaques screened. Mutation frequency is determined as the fraction of cells carrying definitely independent mutations and therefore requires correction for clonal expansion. Mutant and mutation frequencies were determined for brain, thymus and male germ cells of four mice from two age groups (3-versus 10-month old). The mutant frequency in thymus differed significantly between 3- and 10-month old mice (P < 0.05). By sequencing all mutants, the mutation frequency (i.e., corrected for jackpot mutations) in thymus was determined and was not significantly different between 3- and 10-month old mice. Mutant frequency does not fit a Poisson distribution, but mutation frequency corrected for jackpot mutations is substantially less variable and does fit a Poisson distribution.