Quantifying microsatellite mutation rates from intestinal stem cell dynamics in Msh2-deficient murine epithelium
Authors
Morrissey, Edward
Lourenco, Filipe C
Abujudeh, Sam
Thorsen, Anne-Sofie
Hazelwood, Lee
Kemp, Richard
Christopher, Joseph
Potter, Paul K
Publication Date
2019-07Journal Title
Genetics
ISSN
0016-6731
Publisher
Genetics Society of America
Volume
212
Issue
3
Pages
655-665
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Winton, D., Morrissey, E., Lourenco, F. C., Abujudeh, S., Thorsen, A., Hazelwood, L., Kemp, R., et al. (2019). Quantifying microsatellite mutation rates from intestinal stem cell dynamics in Msh2-deficient murine epithelium. Genetics, 212 (3), 655-665. https://doi.org/10.1534/genetics.119.302268
Abstract
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 (around 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 x10-2) compared to wild- type epithelium (6.2 x10-5).
Sponsorship
Cancer Research UK (CB4230)
Wellcome Trust (103805/Z/14/Z)
Embargo Lift Date
2100-01-01
Identifiers
External DOI: https://doi.org/10.1534/genetics.119.302268
This record's URL: https://www.repository.cam.ac.uk/handle/1810/293055
Rights
All rights reserved, Attribution 4.0 International
Licence URL: http://creativecommons.org/licenses/by/4.0/
Statistics
Total file downloads (since January 2020). For more information on metrics see the
IRUS guide.