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Microsatellite sequences have an enhanced susceptibility to mutation, and can act as sentinels indicating elevated mutation rates and increased risk of cancer. The probability of mutant fixation within the intestinal epithelium is dictated by a combination of stem cell dynamics and mutation rate. Here, we exploit this relationship to infer microsatellite mutation rates. First a sensitive, multiplexed, and quantitative method for detecting somatic changes in microsatellite length was developed that allowed the parallel detection of mutant [CA]n sequences from hundreds of low-input tissue samples at up to 14 loci. The method was applied to colonic crypts in Mus musculus, and enabled detection of mutant subclones down to 20% of the cellularity of the crypt (∼50 of 250 cells). By quantifying age-related increases in clone frequencies for multiple loci, microsatellite mutation rates in wild-type and Msh2-deficient epithelium were established. An average 388-fold increase in mutation per mitosis rate was observed in Msh2-deficient epithelium (2.4 × 10-2) compared to wild-type epithelium (6.2 × 10-5).

Original publication

DOI

10.1534/genetics.119.302268

Type

Journal article

Journal

Genetics

Publication Date

07/2019

Volume

212

Pages

655 - 665

Keywords

Msh2, intestine, microsatellite, mouse, mutation, Adult Stem Cells, Animals, Female, Intestinal Mucosa, Male, Mice, Mice, Inbred C57BL, Microsatellite Repeats, Mitosis, MutS Homolog 2 Protein, Mutation Rate