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BOR - Papers in Press, published online ahead of print May 14, 2003.
Biol Reprod 2003, 10.1095/biolreprod.103.017293
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BIOLOGY OF REPRODUCTION 69, 902–914 (2003)
DOI: 10.1095/biolreprod.103.017293
© 2003 by the Society for the Study of Reproduction, Inc.

Reproductive Technology

Disruption of Imprinted Gene Methylation and Expression in Cloned Preimplantation Stage Mouse Embryos1

Mellissa R.W. Mann3, Young Gie Chung4, Leisha D. Nolen3, Raluca I. Verona3, Keith E. Latham2,4, and Marisa S. Bartolomei3

Howard Hughes Medical Institute and Department of Cell and Developmental Biology,3 University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 The Fels Institute for Cancer Research and Molecular Biology and Department of Biochemistry,4 Temple University School of Medicine, Philadelphia, Pennsylvania 19140

Cloning by somatic cell nuclear transfer requires that epigenetic information possessed by the donor nucleus be reprogrammed to an embryonic state. Little is known, however, about this remodeling process, including when it occurs, its efficiency, and how well epigenetic markings characteristic of normal development are maintained. Examining the fate of epigenetic information associated with imprinted genes during clonal development offers one means of addressing these questions. We examined transcript abundance, allele specificity of imprinted gene expression, and parental allele-specific DNA methylation in cloned mouse blastocysts. Striking disruptions were seen in total transcript abundance and allele specificity of expression for five imprinted genes. Only 4% of clones recapitulated a blastocyst mode of expression for all five genes. Cloned embryos also exhibited extensive loss of allele-specific DNA methylation at the imprinting control regions of the H19 and Snprn genes. Thus, epigenetic errors arise very early in clonal development in the majority of embryos, indicating that reprogramming is inefficient and that some epigenetic information may be lost.

1 This research was supported by the National Institutes of Health (HD38381 and 5 T32 CA09214-20) and the Howard Hughes Medical Institute. M.R.W.M. was supported by a grant from The Lalor Foundation. R.I.V. is a Rena and Vic Damone fellow of the American Cancer Society. M.R.W.M, Y.G.C., and L.D.N. contributed equally to this work

2 Correspondence: Keith E. Latham, The Fels Institute for Cancer Research and Molecular Biology and Department of Biochemistry, Temple University School of Medicine, Philadelphia, PA 19140. FAX: 215 707 1454; klatham{at}temple.edu




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