Rational design of artificial ON-riboswitches

Riboswitches are composed of two regions: one for binding to the ligand (the aptamer domain) and the other for regulating the expression of the gene (the expression platform). In most riboswitches (both natural and artificial), a part of the aptamer domain required for ligand binding is directly involved in the regulation of expression, so that it is difficult to design other ligand-responsive riboswitches based on these riboswitches even by using artificial aptamers obtained through in vitro selection. This chapter describes a method for rationally constructing a foundational ON-riboswitch, which is easily available for the design of other ligand-dependent riboswitches, by introducing a new region (a modulator sequence: MS) in addition to the two basic regions. A facile method for preparing arbitrary molecule-dependent riboswitches based on the foundational riboswitch is also presented.