Institute of Inorganic Chemistry, University of Vienna, Josef-Holaubek-Platz 2, 1090 Vienna, Austria.
IBMM, University of Montpellier, CNRS, ENSCM, Montpellier, France.
Sci Adv. 2024 Aug 2;10(31):eado6762. doi: 10.1126/sciadv.ado6762. Epub 2024 Jul 31.
Nucleic acid photolithography is the only microarray fabrication process that has demonstrated chemical versatility accommodating any type of nucleic acid. The current approach to RNA microarray synthesis requires long coupling and photolysis times and suffers from unavoidable degradation postsynthesis. In this study, we developed a series of RNA phosphoramidites with improved chemical and photochemical protection of the 2'- and 5'-OH functions. In so doing, we reduced the coupling time by more than half and the photolysis time by a factor of 4. Sequence libraries that would otherwise take over 6 hours to synthesize can now be prepared in half the time. Degradation is substantially lowered, and concomitantly, hybridization signals can reach over seven times those of the previous state of the art. Under those conditions, high-density RNA microarrays and RNA libraries can now be synthesized at greatly accelerated rates. We also synthesized fluorogenic RNA Mango aptamers on microarrays and investigated the effect of sequence mutations on their fluorogenic properties.
核酸光光刻是唯一一种已经证明具有化学多功能性的微阵列制造工艺，可以适应任何类型的核酸。目前的 RNA 微阵列合成方法需要较长的偶联和光解时间，并且在合成后会不可避免地降解。在这项研究中，我们开发了一系列具有改进的 2'-和 5'-OH 功能的化学和光化学保护的 RNA 亚磷酰胺。这样，我们将偶联时间缩短了一半以上，光解时间缩短了 4 倍。否则需要超过 6 小时才能合成的序列文库现在可以在一半的时间内完成。降解大大降低，同时，杂交信号可以达到以前最先进技术的七倍以上。在这些条件下，高密度的 RNA 微阵列和 RNA 文库现在可以以大大加快的速度合成。我们还在微阵列上合成了荧光 RNA Mango 适体，并研究了序列突变对其荧光性质的影响。
