NT: Abnormal gene expression
Abnormal gene expression in cloned mice derived from embryonic stem cell and cumulus cell nuclei
David Humpherys*,dagger , Kevin Eggan*,dagger , Hidenori AkutsuDagger , Adam Friedman*, Konrad Hochedlinger*, Ryuzo YanagimachiDagger , Eric S. Lander*,dagger , Todd R. Golub*,§,¶, and Rudolf Jaenisch*,dagger ,||
* Whitehead Institute for Biomedical Research and dagger Department of Biology, Massachusetts Institute of Technology, 9 Cambridge Center, Cambridge, MA 02142; Dagger Department of Anatomy and Reproductive Biology, John A. Burns School of Medicine, University of Hawaii, Honolulu, HI 96822; § Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA 02115; and ¶ Department of Pediatrics, Harvard Medical School, Boston, MA 02115
link
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 approx 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.
Fig. 2. (below) 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.
David Humpherys*,dagger , Kevin Eggan*,dagger , Hidenori AkutsuDagger , Adam Friedman*, Konrad Hochedlinger*, Ryuzo YanagimachiDagger , Eric S. Lander*,dagger , Todd R. Golub*,§,¶, and Rudolf Jaenisch*,dagger ,||
* Whitehead Institute for Biomedical Research and dagger Department of Biology, Massachusetts Institute of Technology, 9 Cambridge Center, Cambridge, MA 02142; Dagger Department of Anatomy and Reproductive Biology, John A. Burns School of Medicine, University of Hawaii, Honolulu, HI 96822; § Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA 02115; and ¶ Department of Pediatrics, Harvard Medical School, Boston, MA 02115
link
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 approx 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.
Fig. 2. (below) 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.
posted by nika at 1:49 PM
0 Comments:
Post a Comment
<< Home