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通过酶促转化从 DNA 微阵列到大规模并行 RNA 微阵列的简单合成。

Simple synthesis of massively parallel RNA microarrays via enzymatic conversion from DNA microarrays.

机构信息

Institute of Inorganic Chemistry, University of Vienna, Josef-Holaubek-Platz 2, 1090, Vienna, Austria.

Chair of Food Chemistry and Molecular Sensory Science, Technical University of Munich, Lise-Meitner-Straße 34, 85354, Freising, Germany.

出版信息

Nat Commun. 2022 Jun 30;13(1):3772. doi: 10.1038/s41467-022-31370-9.

Abstract

RNA catalytic and binding interactions with proteins and small molecules are fundamental elements of cellular life processes as well as the basis for RNA therapeutics and molecular engineering. In the absence of quantitative predictive capacity for such bioaffinity interactions, high throughput experimental approaches are needed to sufficiently sample RNA sequence space. Here we report on a simple and highly accessible approach to convert commercially available customized DNA microarrays of any complexity and density to RNA microarrays via a T7 RNA polymerase-mediated extension of photocrosslinked methyl RNA primers and subsequent degradation of the DNA templates.

摘要

RNA 与蛋白质和小分子的催化和结合相互作用是细胞生命过程的基本要素,也是 RNA 治疗和分子工程的基础。在缺乏对这种生物亲和力相互作用进行定量预测的能力的情况下,需要高通量的实验方法来充分采样 RNA 序列空间。在这里,我们报告了一种简单且易于使用的方法,可以通过 T7 RNA 聚合酶介导的光交联甲基 RNA 引物的延伸以及随后的 DNA 模板降解,将任何复杂度和密度的商业上可用的定制 DNA 微阵列转化为 RNA 微阵列。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/54bf/9246885/6ebd324789e3/41467_2022_31370_Fig1_HTML.jpg

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