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Vol. 16, No. 10, pp. 1209-1219, May 15, 2002
1 Germline Development Group, 2 Developmental
Epigenetics Group, Center for Animal Transgenesis and Germ Cell
Research, The School of Veterinary Medicine, University of
Pennsylvania, New Bolton Center, Kennett Square,
Pennsylvania 19348, USA
Somatic cell clones often fail at a developmental stage coincident
with commencement of differentiation. The transcription factor Oct4 is
expressed during cleavage stages and is essential for the
differentiation of the blastocyst. Oct4 expression becomes restricted
to the inner cell mass and epiblast. After gastrulation Oct4 is
active only in germ cells and is silent in somatic cells. Here,
Oct4 and an Oct4-GFP transgene were used as markers
for which gene reprogramming could be directly related to the
developmental potential of somatic cell clones. Cumulus cell clones
initiated Oct4 expression at the correct stage but showed an
incorrect spatial expression in the majority of blastocysts. The
ability of clones to form outgrowths was reduced, and the outgrowths
had low or even undetectable levels of Oct4 RNA or GFP. The quality of
GFP signals in blastocysts correlated with the ability to generate outgrowths that maintain GFP expression and the frequency of embryonic stem (ES) cell derivation. Abnormal Oct4 expression in clones is either directly or indirectly caused by reprogramming errors and is
indicative of a general failure to reset the genetic program. The abnormal Oct4 expression may be associated
with aberrant expression of other crucial developmental genes, leading
to abnormalities at various embryonic stages. Regardless of other
genes, the variations observed in Oct4 levels alone account for the
majority of failures currently observed for somatic cell cloning.
[Key Words: Oct4; pluripotency; Oct4-GFP transgene; gene reprogramming failure; nuclear transfer; blastocyst stage clones]
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