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Background  Animals exhibit circadian rhythms with a period of approximately 24 h in various physiological functions, including locomotor activity. This rhythm is controlled by an endogenous oscillatory mechanism, or circadian clock, which consists of cyclically expressed clock genes and their product proteins.  cryptochrome  ( cry ) genes are thought to be involved in the clock mechanism, and their functions have been examined extensively in holometabolous insects, but in hemimetabolous insects their role is less well understood.    Results  In the present study, the role of  cry  genes was investigated using RNAi technology in a hemimetabolous insect, the cricket  Gryllus bimaculatus . Using a molecular cloning approach, we obtained cDNAs for two  cry  genes:  Drosophila -type  cry1  ( Gb’cry1 ) and mammalian-type  cry2  ( Gb’cry2 ).  Gb’cry2  has six splicing variants, most of which showed rhythmic mRNA expression.  Gb’cry1   RNAi  treatment had only a limited effect at the behavioral and molecular levels, while  Gb’cry2   RNAi  had a significant effect on behavioral rhythms and molecular oscillatory machinery, alone or in combination with  Gb’cry1   RNAi . In  Gb’cry1 / Gb’cry2  double-RNAi crickets, most clock genes showed arrhythmic expression, except for  timeless , which retained clear rhythmic expression. Molecular analysis revealed that some combination of  Gb’cry1  and  Gb’cry2  variants suppressed CLK/CYC transcriptional activity in cultured cells.    Conclusion  Based on these results, we propose a new model of the cricket’s circadian clock, including a molecular oscillatory loop for  Gb’cry2 , which can operate independent of the  Gb’per / Gb’tim  loop.

cryptochrome genes form an oscillatory loop independent of the per/tim loop in the circadian clockwork of the cricket Gryllus bimaculatus