title: RNA-based fluorescent biosensors for live cell imaging of second messengers cyclic di-GMP and cyclic AMP-GMP.
abstract: 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.
authors.affiliation: Department of Chemistry and ‡Department of Molecular & Cell Biology, University of California , Berkeley, California 94720, United States.
authors.forename: Colleen A
authors.initials: CA
authors.lastname: Kellenberger
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Wilson
authors.forename: Jade
authors.initials: J
authors.lastname: Sales-Lee
authors.forename: Ming C
authors.initials: MC
authors.lastname: Hammond
mesh_terms: D000595:Amino Acid Sequence; D052157:Aptamers, Nucleotide / Q000235:genetics; D015374:Biosensing Techniques / Q000379:methods*; D000242:Cyclic AMP / Q000737:chemistry* / Q000235:genetics; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000235:genetics; D005453:Fluorescence; D057054:Molecular Imaging / Q000379:methods*; D012313:RNA / Q000737:chemistry*; D018724:Spinacia oleracea / Q000235:genetics
chemical_list: D052157:Aptamers, Nucleotide; C062025:bis(3',5')-cyclic diguanylic acid; D012313:RNA; D000242:Cyclic AMP; D006152:Cyclic GMP