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. 2002 Oct 1;99(20):12889-94.
doi: 10.1073/pnas.192433399. Epub 2002 Sep 16.

Abnormal gene expression in cloned mice derived from embryonic stem cell and cumulus cell nuclei

David Humpherys  1 , Kevin Eggan, Hidenori Akutsu, Adam Friedman, Konrad Hochedlinger, Ryuzo Yanagimachi, Eric S Lander, Todd R Golub, Rudolf Jaenisch
Affiliations

Abnormal gene expression in cloned mice derived from embryonic stem cell and cumulus cell nuclei

David Humpherys et al. Proc Natl Acad Sci U S A. .

Abstract

To assess the extent of abnormal gene expression in clones, we assessed global gene expression by microarray analysis on RNA from the placentas and livers of neonatal cloned mice derived by nuclear transfer (NT) from both cultured embryonic stem cells and freshly isolated cumulus cells. Direct comparison of gene expression profiles of more than 10,000 genes showed that for both donor cell types approximately 4% of the expressed genes in the NT placentas differed dramatically in expression levels from those in controls and that the majority of abnormally expressed genes were common to both types of clones. Importantly, however, the expression of a smaller set of genes differed between the embryonic stem cell- and cumulus cell-derived clones. The livers of the cloned mice also showed abnormal gene expression, although to a lesser extent, and with a different set of affected genes, than seen in the placentas. Our results demonstrate frequent abnormal gene expression in clones, in which most expression abnormalities appear common to the NT procedure whereas others appear to reflect the particular donor nucleus.

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Figures

Figure 1
Figure 1
(A) Representation of the expression levels of all genes showing a 2-fold difference in mean expression between any two sample types. Placentas are along the vertical axis and genes along the horizontal axis. Average expression across the 15 samples is indicated by black. Expression more than the average level is indicated by increasing red intensity, and green indicates reduced expression. Genes with similar expression profiles have been clustered and displayed with genecluster and treeview. (B) The number of genes that show a 2-fold difference in average expression and differing significantly from controls by a t test (P < 0.05) are indicated for each of the donor cell types beneath the diagrams. There are two separated diagrams for genes either reduced or elevated in the NT placentas. The number of genes altered 2-fold in both types of clones is indicated in the overlapping circles.
Figure 2
Figure 2
Northern analysis of several genes dysregulated in NT placentas at term. Lanes 1–6 contain RNA from naturally derived B6/129 controls, whereas the RNAs in lanes 7 and 8 are derived from the placentas from normal B6/129 zygotes that had been cultured in vitro before transfer to a surrogate mother. RNAs in lanes 9–18 are from cumulus cell NT placentas of the indicated genetic backgrounds: lanes 9–13, DBA/Cast; lanes 14 and 15, 129/Cast; lane 16, AJ/Cast, and lanes 17 and 18, B6/DBA. RNAs in lanes 19–37 are from placentas of ES cell NT mice: lanes 19–29 are derived from the V6.5 (B6/129) line, lanes 30–33 are from targeted subclones (14) of the V6.5 line (lanes 30–32, subclone 89; lane 33, subclone 23), lane 34 is from the V17.2 (BALB/129) line, and lanes 35–37 are from the F1–2.3 (129/Cast) line.
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References

    1. Young L E, Sinclair K D, Wilmut I. Rev Reprod. 1998;3:155–163. - PubMed
    1. Chavatte-Palmer P, Heyman Y, Renard J P. Gynecol Obstet Fertil. 2000;28:633–642. - PubMed
    1. Wakayama T, Yanagimachi R. Nat Genet. 1999;22:127–128. - PubMed
    1. Hill J R, Roussel A J, Cibelli J B, Edwards J F, Hooper N L, Miller M W, Thompson J A, Looney C R, Westhusin M E, Robl J M, Stice S L. Theriogenology. 1999;51:1451–1465. - PubMed
    1. Hill J R, Burghardt R C, Jones K, Long C R, Looney C R, Shin T, Spencer T E, Thompson J A, Winger Q A, Westhusin M E. Biol Reprod. 2000;63:1787–1794. - PubMed

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