In Vivo Structure-Activity Relationships and Optimization of an Unnatural Base Pair for Replication in a Semi-Synthetic Organism
In Vivo Structure-Activity Relationships and Optimization of an Unnatural Base Pair for Replication in a Semi-Synthetic Organism
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DOI：
10.1021/jacs.7b03540
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发表时间：
2017-08-23
影响因子：
15
通讯作者：
Romesberg, Floyd E.
中科院分区：
文献类型：
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作者：
Feldman, Aaron W.;Romesberg, Floyd E.
为了扩展遗传字母表并创建存储和检索更多信息的半合成生物体（SSO），我们开发了非天然碱基对（UBP）dNaM和dSSICS或dTPT 3（dNaM-dSSICS和dNaM-dTPT 3）。UBPs的形成基于疏水力和填充力，而不是互补氢键，虽然它们都保留在大肠杆菌SSO的体内环境中，但它们的发展是基于体外产生的结构-活性关系（SAR）数据。为了解决体内环境不同要求的可能性，我们筛选了135个候选UBP，以获得SSO中的最佳性能。有趣的是，我们发现体内SAR不同于体外收集的SAR，最重要的是，我们确定了四种UBP，其在SSO DNA中的保留高于dNaM-dTPT 3，这是以前确定的最有前途的UBP。这四个UBP的鉴定进一步表明，当优化时，疏水和包装力可以用来取代天然对所使用的互补氢键，并代表了我们在开发存储和检索比天然生物更多信息的SSO的持续努力中的重大进展。
In an effort to expand the genetic alphabet and create semi-synthetic organisms (SSOs) that store and retrieve increased information, we have developed the unnatural base pairs (UBPs) dNaM and dSSICS or dTPT3 (dNaM-dSSICS and dNaM-dTPT3). The UBPs form based on hydrophobic and packing forces, as opposed to complementary hydrogen bonding, and while they are both retained within the in vivo environment of an Escherichia coli SSO, their development was based on structure-activity relationship (SAR) data generated in vitro. To address the likely possibility of different requirements of the in vivo environment, we screened 135 candidate UBPs for optimal performance in the SSO. Interestingly, we find that in vivo SARs differ from those collected in vitro, and most importantly, we identify four UBPs whose retention in the DNA of the SSO is higher than that of dNaM-dTPT3, which was previously the most promising UBP identified. The identification of these four UBPs further demonstrates that when optimized, hydrophobic and packing forces may be used to replace the complementary hydrogen bonding used by natural pairs and represents a significant advance in our continuing efforts to develop SSOs that store and retrieve more information than natural organisms.