Progress toward Eukaryotic Semisynthetic Organisms: Translation of Unnatural Codons.
Progress toward Eukaryotic Semisynthetic Organisms: Translation of Unnatural Codons.
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DOI：
10.1021/jacs.9b09080
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发表时间：
2019-12
影响因子：
15
通讯作者：
Anne Xiao-Zhou Zhou-Anne-Xiao-Zhou-Zhou-13769862;Kai Sheng;Aaron W Feldman;F. Romesberg
中科院分区：
文献类型：
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作者：
Anne Xiao-Zhou Zhou-Anne-Xiao-Zhou-Zhou-13769862;Kai Sheng;Aaron W Feldman;F. Romesberg
我们已经创建了一个细菌半合成生物体（SSO），它在其DNA中保留了一个非天然碱基对（UBP），将其转录为具有同源非天然密码子和反密码子的mRNA和tRNA，并在tRNA上加载非规范氨基酸后合成含有非规范氨基酸的蛋白质。在这里，我们报告的第一个真核SSO的创建进展。在证明了与人HEK 293细胞的概念验证后，我们发现各种不同的非天然密码子-反密码子对可以有效地介导CHO细胞中非天然蛋白质的合成。有趣的是，我们发现原核和真核核糖体识别UBP的方式既有相似之处，也有显着差异，真核核糖体表现出更大的耐受性。这些结果代表了真核SSO的第一个进展，事实上，这表明这样的SSO可能能够保留比细菌更多的非自然信息。
We have created a bacterial semisynthetic organism (SSO) that retains an unnatural base pair (UBP) in its DNA, transcribes it into mRNA and tRNA with cognate unnatural codons and anticodons, and after the tRNA is charged with a noncanonical amino acid synthesizes proteins containing the noncanonical amino acid. Here, we report the first progress toward the creation of eukaryotic SSOs. After demonstrating proof-of-concept with human HEK293 cells, we show that a variety of different unnatural codon-anticodon pairs can efficiently mediate the synthesis of unnatural proteins in CHO cells. Interestingly, we find that there are both similarities and significant differences between how the prokaryotic and eukaryotic ribosomes recognize the UBP, with the eukaryotic ribosome appearing more tolerant. The results represent the first progress toward eukaryotic SSOs and, in fact, suggest that such SSOs might be able to retain more unnatural information than their bacterial counterparts.