Department of Pharmaceutical Sciences, University of California, Irvine, Irvine, California 92697, United States.
Universidad Europea de Madrid, E-28670 Villaviciosa de Odon, Madrid Spain.
ACS Chem Biol. 2020 Dec 18;15(12):3099-3105. doi: 10.1021/acschembio.0c00755. Epub 2020 Nov 21.
Profiling RNA expression in a cell-specific manner continues to be a grand challenge in biochemical research. Bioorthogonal nucleosides can be utilized to track RNA expression; however, these methods currently have limitations due to background and incorporation of analogs into undesired cells. Herein, we design and demonstrate that uracil phosphoribosyltransferase can be engineered to match 5-vinyluracil for cell-specific metabolic labeling of RNA with exceptional specificity and stringency.
以细胞特异性方式进行 RNA 表达谱分析仍然是生化研究中的一个重大挑战。生物正交核苷可用于跟踪 RNA 表达；然而，由于背景和类似物掺入非预期细胞，这些方法目前存在局限性。在此，我们设计并证明可以对尿嘧啶磷酸核糖基转移酶进行工程改造，使其与 5-乙烯基尿嘧啶匹配，从而实现 RNA 的细胞特异性代谢标记，具有出色的特异性和严格性。
