In vivo mutation analysis using the ΦX174 transgenic mouse and comparisons with other transgenes and endogenous genes.
The ΦX174 transgenic mouse was first developed as an in vivo Ames test, detecting base pair substitution (bps) at a single bp in a reversion assay. A forward mutational assay was also developed, which is a gain of function assay that also detects bps exclusively. Later work with both assays focused on establishing that a mutation was fixed in vivo using single-burst analysis: determining the number of mutant progeny virus from an electroporated cell by dividing the culture into aliquots before scoring mutants. We review results obtained from single-burst analysis, including testing the hypothesis that high mutant frequencies (MFs) of G:C to A:T mutation recovered by transgenic targets include significant numbers of unrepaired G:T mismatches. Comparison between the ΦX174 and lacI transgenes in mouse spleen indicates that the spontaneous bps mutation frequency per nucleotide (mf(n)) is not significantly lower for ΦX174 than for lacI; the response to ENU is also comparable. For the lacI transgene, the spontaneous bps mf(n) is highly age-dependent up to 12 weeks of age and the linear trend extrapolates at conception to a frequency close to the human bps mf(n) per generation of 1.7 × 10(-8). Unexpectedly, we found that the lacI somatic (spleen) bps mf(n) per cell division at early ages was estimated to be the same as for the human germ-line. The bps mf(n) in bone marrow for the gpt transgene is comparable to spleen for the lacI and ΦX174 transgenes. We conclude that the G:C to A:T transition is characteristic of spontaneous in vivo mutation and that the MFs measured in these transgenes at early ages reflect the expected accumulation of in vivo mutation typical of endogenous mammalian mutation rates. However, spontaneous and induced mf(n)s per nucleotide for the cII gene in spleen are 5-10 times higher than for these other transgenes.
Valentine
Carrie R
CR
Division of Genetic and Reproductive Toxicology, National Center for Toxicological Research, 3900 NCTR Road, HFT-120, Jefferson, AR 72079, USA.
Delongchamp
Robert R
RR
Pearce
Mason G
MG
Rainey
Heather F
HF
Dobrovolsky
Vasily N
VN
Malling
Heinrich V
HV
Heflich
Robert H
RH
D000818:Animals; D010584:Bacteriophage phi X 174 / Q000235:genetics*; D004252:DNA Mutational Analysis; D005821:Genetic Techniques; D005854:Germ Cells / Q000166:cytology; D006801:Humans; D056968:Lac Repressors / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D008957:Models, Genetic; D009154:Mutation; D013154:Spleen / Q000378:metabolism; D013997:Time Factors; D019076:Transgenes
D056968:Lac Repressors