title: In vitro selection of allosteric ribozymes that sense the bacterial second messenger c-di-GMP.
abstract: Recently, a number of study have shown the ligand-dependent allosteric ribozymes can be harnessed as biosensors, high-throughput screening, and agents for the control of gene expression in vivo, called artificial riboswitches. In this chapter, we describe how in vitro selection can be used to create an allosteric ribozyme that senses bacterial second messenger cyclic-di-GMP (c-di-GMP). A hammerhead ribozyme was joined to a natural c-di-GMP class I riboswitch aptamer via communication modules. Both c-di-GMP-activating and -inhibiting ribozyme can be obtained by this approach.
authors.affiliation: Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT, USA.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Furukawa
authors.forename: Hongzhou
authors.initials: H
authors.lastname: Gu
authors.forename: Ronald R
authors.initials: RR
authors.lastname: Breaker
mesh_terms: D000494:Allosteric Regulation; D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D058928:Riboswitch
chemical_list: D052157:Aptamers, Nucleotide; D016337:RNA, Catalytic; D058928:Riboswitch; C427204:hammerhead ribozyme; C062025:bis(3',5')-cyclic diguanylic acid; D006152:Cyclic GMP