Emergency prevention of extinction of a transgenic allele in a less-fertile transgenic mouse line by crossing with an inbred or outbred mouse strain coupled with assisted reproductive technologies

被引:2
作者
Mayer, Anna
Bulian, Diana
Scherb, Hagen
de Angelis, Martin Hrabe
Schmidt, Joerg
Mahabir, Esther [1 ]
机构
[1] GSF, Natl Res Ctr Environm & Hlth, Dept Comparat Med, D-85764 Neuherberg, Germany
[2] GSF, Natl Res Ctr Environm & Hlth, Inst Biomath & Biometry, D-85764 Neuherberg, Germany
[3] GSF, Natl Res Ctr Environm & Hlth, Inst Expt Genet, D-85764 Neuherberg, Germany
关键词
D O I
10.1071/RD06161
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Certain transgenic mouse lines are difficult to breed or archive and, consequently, their transgenes become lost. We examined a C57BL/6 mouse line (B6-tg), transgenic for green fluorescent protein (GFP) with low fertility, and its crosses with the more prolific inbred C3HeB/FeJ (C3) and outbred Swiss (SW) strains in order to assess the possibility of emergency prevention of extinction of a transgenic allele by using assisted reproductive technologies (ART). Out-crossing was performed by natural mating or in vitro fertilisation (IVF) with heterozygous mice. Most of the crossing combinations resulted in improved archiving and rederivation efficiencies of the transgenic allele. Natural crossing increased both mean litter size by two to three pups and the superovulatory rate from 69% for B6-tg to 70-90% for females from the out-crosses. Each plug-positive B6-tg female yielded an average of 4.6 two-cell embryos, whereas females from the out-crosses produced three-to fivefold that amount. After thawing, 13% of B6-tg embryos and 6-12% of out-crossed embryos developed into transgenic pups after transfer into recipients. After IVF with cryopreserved spermatozoa, cleavage rates were 4% for B6-tg, 22-37% for B6-tg oocytes out-crossed with C3 and SW spermatozoa, 9-49% for gametes from out-crossed mice and 28-44% for back-crosses with B6 oocytes. Transgenic pups were not derived from IVF with B6-tg gametes when either fresh or cryopreserved spermatozoa were used. Rederivation efficiencies were 7% and 4% from out-crosses of B6-tg oocytes with C3 and SW spermatozoa, respectively, 6-22% for gametes from out-crossed mice and 4-10% for the back-crosses. Although out-crossing changes the original genetic background, the strategy of crossing coupled with ART prevents the extinction of an allele of interest, especially where archiving and rederivation of the transgenic line fail.
引用
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页码:984 / 994
页数:11
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