Department of Chemistry and ‡Department of Molecular & Cell Biology, University of California , Berkeley, California 94720, United States.
J Am Chem Soc. 2013 Apr 3;135(13):4906-9. doi: 10.1021/ja311960g. Epub 2013 Mar 21.
Cyclic dinucleotides are an important class of signaling molecules that regulate a wide variety of pathogenic responses in bacteria, but tools for monitoring their regulation in vivo are lacking. We have designed RNA-based fluorescent biosensors for cyclic di-GMP and cyclic AMP-GMP by fusing the Spinach aptamer to variants of a natural GEMM-I riboswitch. In live cell imaging experiments, these biosensors demonstrate fluorescence turn-on in response to cyclic dinucleotides, and they were used to confirm in vivo production of cyclic AMP-GMP by the enzyme DncV.
环二核苷酸是一类重要的信号分子，可调节细菌中多种致病反应，但缺乏监测其体内调节的工具。我们通过将 Spinach 适体融合到天然 GEMM-I 核糖开关的变体上，设计了用于检测环二鸟苷酸 (cyclic di-GMP) 和环腺苷酸-鸟苷酸 (cyclic AMP-GMP) 的基于 RNA 的荧光生物传感器。在活细胞成像实验中，这些生物传感器在响应环二核苷酸时表现出荧光开启，并用它们证实了酶 DncV 在体内产生环 AMP-GMP。
