Gene activation-associated long noncoding RNAs function in mouse preimplantation development.
Bottom Line: Expression of these bidirectional promoter-associated noncoding RNAs (pancRNAs) was strongly associated with the upregulation of their cognate genes.Conversely, knockdown of three abundant pancRNAs led to reduced mRNA expression, accompanied by sustained DNA methylation even in the presence of enzymes responsible for DNA demethylation.Thus, this novel class of lncRNAs can modulate the transcription machinery in cis to activate zygotic genes and is important for preimplantation development.
Affiliation: Department of Biophysics and Global COE Program, Graduate School of Science, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto 606-8502, Japan Division of Basic Stem Cell Biology, Department of Stem Cell Biology and Medicine, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.Show MeSH
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Mentions: Since the effect of pancIl17d knockdown was drastic, we focused on investigating the roles of this pancRNA in embryonic development. Many pancIl17d knockdown embryos died between the 8-cell and early blastocyst stages. To establish whether cell death was enhanced in the pancIl17d knockdown embryos, we performed TdT-mediated dUTP nick-end labeling (TUNEL) staining of the pancIl17d knockdown embryos at the morula stage (Fig. 4A). Consistent with a previous report (Brison and Schultz, 1997), control embryos underwent little apoptosis during blastocyst formation. By contrast, pancIl17d knockdown embryos exhibited multiple TUNEL-positive blastomeres, suggesting that many pancIl17d knockdown embryos died by the blastocyst stage due to excessive apoptosis. It is noteworthy that the developmental capacity to form a blastocyst was restored when recombinant mouse IL17D protein (rIL17D) was added to the medium at the 4-cell stage, although the knockdown effect continued until the morula stage (Fig. 4B,C). The addition of rIL17D significantly increased the rate of success of blastocyst formation in pancIl17d knockdown embryos (from 21.5±2.7% to 62.1±5.9%; Fig. 4D). These results suggest that pancIl17d plays an important role in blastocyst formation by upregulating the partner gene.Fig. 4.
Affiliation: Department of Biophysics and Global COE Program, Graduate School of Science, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto 606-8502, Japan Division of Basic Stem Cell Biology, Department of Stem Cell Biology and Medicine, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.