Mechanisms of maternal mRNA-specific knockdown in Ciona intestinalis

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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.