Sequence-based analysis of mutagenized mice.
Treating mice with ethylnitrosourea (ENU) is an efficient means for mutagenizing spermatogonial cells, and this treatment has been proven effective in a variety of screens for both dominant and recessive mutations. However, a significant problem for this technology is that the efficiency of mutagenesis is assessed most often by the empiric determination of a per-locus mutation frequency by using the specific locus test, which is expensive, time-consuming, and logistically difficult. To approach this question more directly and more efficiently, one can utilize methods of PCR-based mutation detection for the characterization of progeny of mutagenized mice. Since this analysis can be done after a single generation of breeding, it is useful as a rapid means for the assessment of the efficiency of mutagen treatment. Furthermore, it is readily imaginable that this strategy can be applied for the general determination of gene function in a systematic manner. Theoretical considerations and empirical analysis suggest that the per-base mutation frequency for a fractionated-dose treatment protocol is on the order of 1 sequence change per 10(5) bp.
Beier
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Genetics Division, Brigham and Women's Hospital, 20 Shattuck Street, Boston, Massachusetts 02115, USA. beier@rascal.med.harvard.edu
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004252:DNA Mutational Analysis; D005038:Ethylnitrosourea; D006580:Genetic Carrier Screening; D005820:Genetic Testing; D051379:Mice / Q000235:genetics*; D016296:Mutagenesis; D009153:Mutagens; D018807:Polymorphism, Single-Stranded Conformational
D009153:Mutagens; D005038:Ethylnitrosourea