Transcriptome analysis after disruption of nuclear F-actin in mouse embryos

We find that filamentous actin (F-actin) is formed in pronuclei of mouse zygotes, disruption of which resulted in abnormal development of mouse embryos. To further explain the molecular mechanism, transcriptome analysis was performed following the artificial disruption of nuclear F-actin by overexpression of mutant actin R62D or G15S in nuclei. Overall design: Nuclear F-actin in zygotes was disrupted by overexpressing actin mutant R62D in pronuclei (R62D-1 and R62D-2). As controls, wild type actin was overexpressed in nuclei (WT-1 or WT-2) or non-injected embryos were used (Actin-control-1 and Actin-control-2). In a different set of experiments, Nuclear F-actin in zygotes was disrupted by overexpressing Exportin 6 together with actin mutant R62D in pronuclei (Exp6+R62D-1-3) or abnormal nuclear F-actin formation was induced by expressing actin mutant G15S in pronuclei (G15S-1-3). As a control, non-injected embryos were used (Non-1-3). RNA-seq was performed using the mouse 2-cell stage embryos, collected at 32 hours post insemination.