title: Improvement of ENU Mutagenesis Efficiency Using Serial Injection and Mismatch Repair Deficiency Mice.
abstract: ENU mutagenesis is a powerful method for generating novel lines of mice that are informative with respect to both fundamental biological processes and human disease. Rapid developments in genomic technology have made the task of identifying causal mutations by positional cloning remarkably efficient. One limitation of this approach remains the mutation frequency achievable using standard treatment protocols, which currently generate approximately 1-2 sequence changes per megabase when optimized. In this study we used two strategies to attempt to increase the number of mutations induced by ENU treatment. One approach employed mice carrying a mutation in the DNA repair enzyme Msh6. The second strategy involved injection of ENU to successive generations of mice. To evaluate the number of ENU-induced mutations, single mice or pooled samples were analyzed using whole exome sequencing. The results showed that there is considerable variability in the induced mutation frequency using these approaches, but an overall increase in ENU-induced variants from one generation to another was observed. The analysis of the mice deficient for Msh6 also showed an increase in the ENU-induced variants compared to the wild-type ENU-treated mice. However, in both cases the increase in ENU-induced mutation frequency was modest.
authors.affiliation: Center for Developmental Biology and Regenerative Medicine, Seattle Children's Research Institute, Seattle, WA, United States of America.|Department of Pediatrics, University of Washington School of Medicine, Seattle, WA, United States of America.
authors.forename: Jabier
authors.initials: J
authors.lastname: Gallego-Llamas
authors.affiliation: Center for Developmental Biology and Regenerative Medicine, Seattle Children's Research Institute, Seattle, WA, United States of America.
authors.forename: Andrew E
authors.initials: AE
authors.lastname: Timms
authors.affiliation: McLaughlin Research Institute, Great Falls, MT, United States of America.
authors.forename: Rose
authors.initials: R
authors.lastname: Pitstick
authors.affiliation: McLaughlin Research Institute, Great Falls, MT, United States of America.
authors.forename: Janet
authors.initials: J
authors.lastname: Peters
authors.affiliation: McLaughlin Research Institute, Great Falls, MT, United States of America.
authors.forename: George A
authors.initials: GA
authors.lastname: Carlson
authors.affiliation: Center for Developmental Biology and Regenerative Medicine, Seattle Children's Research Institute, Seattle, WA, United States of America.|Department of Pediatrics, University of Washington School of Medicine, Seattle, WA, United States of America.
authors.forename: David R
authors.identifier: http://orcid.org/0000-0002-0325-8520
authors.initials: DR
authors.lastname: Beier
mesh_terms: D000818:Animals; D020029:Base Pairing / Q000235:genetics; D001947:Breeding; D053843:DNA Mismatch Repair / Q000235:genetics*; D005038:Ethylnitrosourea / Q000737:chemistry*; D005260:Female; D005298:Fertility; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D016296:Mutagenesis / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D016019:Survival Analysis
chemical_list: D005038:Ethylnitrosourea