Abstract
Cloned pigs generated by somatic cell nuclear transfer (SCNT) show a greater ratio of early abortion during mid-gestation than normal controls. X-linked genes have been demonstrated to be important for the development of cloned embryos. To determine the relationship between the expression of X-linked genes and abortion of cloned porcine fetuses, the expression of X-linked genes were investigated by quantitative real-time polymerase chain reaction (q-PCR) and the methylation status of Xist DMR was performed by bisulfate-specific PCR (BSP). q-PCR analysis indicated that there was aberrant expression of X-linked genes, especially the upregulated expression of Xist in both female and male aborted fetuses compared to control fetuses. Results of BSP suggested that hypomethylation of Xist occurred in aborted fetuses, whether male or female. These results suggest that the abnormal expression of Xist may be associated with the abortion of fetuses derived from somatic cell nuclear transfer embryos.
Highlights
Since the first cloned piglets were generated in 2000, a large number of genetically modified somatic cell nuclear transfer (SCNT) cloned pigs have been harvested in a number of laboratories worldwide [1,2,3]
Considering that there was possibly gene expression discrepancy between male and female aborted fetus, sex identification was performed with genomic DNA from the aborted fetuses and SCNT donor cells
The expression of ARAF1 was higher in male aborted fetuses than in male control fetuses, and there was no significant difference between female control and aborted fetuses (Figure 2C). These results indicate that proper regulation of the expression of X inactivation-specific transcript (Xist) may be critical for the development of cloned fetuses
Summary
Since the first cloned piglets were generated in 2000, a large number of genetically modified SCNT cloned pigs have been harvested in a number of laboratories worldwide [1,2,3]. SCNT is a useful technique for embryos production in many fields, such as animal models, xenotransplantation and embryonic stem cell research. Many genetically modified cloned pigs have been generated, there are still problems that must be resolved. The general cloning efficiency remains unsatisfactory, at lower than 1.5%, it is improving [4]. A certain proportion of surrogate sows for transferred SCNT embryos suffer abortions during gestation, and these may be concentrated between Day 27 and Day 34 following SCNT [5]. Some cloned piglets suffer from serious developmental abnormalities and die within a few days of birth
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