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TRIM28/KAP1/TIF1β was identified as a universal transcriptional co-repressor and is critical for regulating post-fertilization methylation reprogramming in preimplantation embryos. In this study, three siRNAs (si647, si742, and si1153) were designed to target the TRIM28 mRNA sequence. After transfection of the mixture of the three siRNA (siMix) into bovine fibroblast cells, the most effective one for TRIM28 knockdown was selected. By injecting RNAi directed against TRIM28 mRNA, we found that TRIM28 knockdown in oocytes had the most effect on the H19 gene, in which differentially methylated region (DMR) methylation was almost completely absent at the 2-cell stage (1.4%), while control embryos showed 74% methylation. In addition, global H3K9me3 levels at the 2-cell stage were significantly higher in the in vitro fertilization (IVF) group than in the TRIM28 knockdown group (P<0.05). We further show that TRIM28 is highly expressed during oocyte maturation and reaches peak levels at the 2-cell stage. In contrast, at this stage, TRIM28 expression in somatic cell nuclear transfer (SCNT) embryos decreased significantly (P<0.05), suggesting that Trim28 transcripts are lost during SCNT. TRIM28 is required for the maintenance of methylation imprints in bovine preimplantation embryos, and the loss of TRIM28 during SCNT may contribute to the unfaithful maintenance of imprints in cloned embryos.
A 50-yr resolution reconstruction of climate and environment variability during the period 43–14 ka was developed using 26 high-precision U/Th dates and 390 oxygen isotope (δ18O) data of a stalagmite (SJ1) collected from Songjia Cave in central China, which is close to the northwestern boundary of the Asian summer monsoon (ASM). The δ18O record in SJ1 displays significant millennial-scale changes that correlate well in timing and duration with Dansgaard/Oeschger (D/O) events 5–10 and Heinrich event 4 (H4) identified in high-latitude regions of the Northern Hemisphere. Four 230Th dates constrain the H4 event precisely to the period of 39.7 to 38.3 ka. Notable centennial variations of the ASM activity could be observed within the H4 event. The magnitude and duration of D/O event 4.1 recorded in SJ1 are similar to those archived in east China but different from those documented in southwest China, suggesting that the manifestation of this event may be regionally different. The timing, duration and structure of D/O events 5–10 and Heinrich event 4 suggest that temperature changes in both hemispheres have exerted significant influences on the ASM variations in central China.
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