Mechanisms of maternal mRNA-specific knockdown in Ciona intestinalis

本研究旨在对海鞘Ciona mRNAs的靶向和特异性敲除方法进行表征，并了解敲除方法对海鞘Ciona mRNAs的定位机制。我们确定了基因敲除所需的DNA元件。eGFP、Kaede、mKO 2和DsRed报告基因在与靶基因的启动子和5' UTR一起表达时，可引起靶基因的敲低。Minos和睡美人转座子都可以用于敲低方法。我们发现5 'UTR对于确定敲除的靶基因是至关重要的，并且我们表明在敲除系中从5' UTR产生小的反义RNA。我们敲除了编码Y盒型RNA结合蛋白的母源表达基因Ci-YB 1。敲除母体Ci-YB 1的动物显示出异常发育，表明该基因是正常胚胎发育所必需的。

This research purposed the characterization of the targeted and specific knockdown method of maternal mRNAs in the ascidian Ciona intestinalis, and understanding the mechanisms of the localization of maternal mRNAs in Ciona with the knockdown method. We identified the DNA elements necessary for the knockdown. eGFP, Kaede, mKO2 and DsRed reporter genes can cause knockdown of target genes when expressed with the promoter and 5' UTR of target genes. Both Minos and Sleeping Beauty transposons can be used for the knockdown method. We found that 5'UTR is crucial for determining the target gene of knockout, and we showed that small antisense RNAs are produced from the 5'UTR in the knockdown lines. We knocked down a maternally expressed gene Ci-YB1 that encodes Y-box type RNA-binding protein. The animals in which maternal Ci-YB1 was knocked down showed abnormal development, suggesting that this gene is necessary for proper embryogenesis.