高密度RNA阵列的酶促制备。
Enzymatic fabrication of high-density RNA arrays.
作者信息
Wu Cheng-Hsien, Holden Matthew T, Smith Lloyd M
机构信息
Department of Chemistry, University of Wisconsin at Madison, 1101 University Avenue, Madison, WI 53706 (USA).
出版信息
Angew Chem Int Ed Engl. 2014 Dec 1;53(49):13514-7. doi: 10.1002/anie.201408747. Epub 2014 Oct 22.
Abstract
A powerful new strategy for the fabrication of high-density RNA arrays is described. A high-density DNA array is fabricated by standard photolithographic methods, the surface-bound DNA molecules are enzymatically copied into their RNA complements from a surface-bound RNA primer, and the DNA templates are enzymatically destroyed, leaving behind the desired RNA array. The strategy is compatible with 2'-fluoro-modified (2'F) ribonucleoside triphosphates (rNTPs), which may be included in the polymerase extension reaction to impart nuclease resistance and other desirable characteristics to the synthesized RNAs. The use and fidelity of the arrays are explored with DNA hybridization, DNAzyme cleavage, and nuclease digestion experiments.
摘要
描述了一种用于制造高密度RNA阵列的强大新策略。通过标准光刻方法制造高密度DNA阵列，将表面结合的DNA分子从表面结合的RNA引物酶促复制成其RNA互补物，然后酶促破坏DNA模板，留下所需的RNA阵列。该策略与2'-氟修饰（2'F）的核糖核苷三磷酸（rNTP）兼容，其可包含在聚合酶延伸反应中，以使合成的RNA具有核酸酶抗性和其他所需特性。通过DNA杂交、DNA酶切割和核酸酶消化实验探索了阵列的用途和保真度。
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