Generation of transgenic rabbits by the novel technique of chimeric somatic cell cloning

Biology of Reproduction
M SkrzyszowskaM Pieńkowski

Abstract

A novel technique of chimeric somatic cell cloning was applied to produce a transgenic rabbit (NT20). Karyoplasts of transgenic adult skin fibroblasts with Tg(Wap-GH1) gene construct as a marker were microsurgically transferred into one, previously enucleated, blastomere of 2-cell non-transgenic embryos, while the second one remained intact. The reconstructed embryos either were cultured in vitro up to the blastocyst stage (Experiment I) or were transferred into recipient-females immediately after the cloning procedure (Experiment II). In Experiment I, 25/102 (24.5%) embryos formed blastocysts from whole embryos and 46/102 (44.12%) embryos developed to the blastocyst stage from single non-operated blastomeres, while the reconstructed blastomeres were damaged and degenerated. Thirteen (12.7%) embryos did not exceed 3- to 4-cell stages and 18 (17.7%) embryos were inhibited at the initial 2-cell stage. Out of 14 blastocysts which were subjected to molecular analysis, the transgene was detected in the cells of 4 blastocysts. In Experiment II, 163/217 (75.0%) embryos were transferred into 9 pseudopregnant recipient-rabbits (an average of 18 embryos per recipient). Four recipient-females (44.4%) became pregnant and delivered a total ...Continue Reading

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Citations

Oct 31, 2006·Reproduction : the Official Journal of the Society for the Study of Fertility·Pawel GredaJacek A Modlinski
May 31, 2016·FEBS Open Bio·Yongye HuangDaxin Pang
Feb 20, 2021·Frontiers in Genetics·Jianglin FanY Eugene Chen

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Methods Mentioned

BETA
transgenic
biopsy
PCR
biopsies
genotyping

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