CLOCK evolved in cnidaria to synchronize internal rhythms with diel environmental cues

The circadian clock enables anticipation of the day/night cycle in animals ranging from cnidarians to mammals. Circadian rhythms are generated through a transcription-translation feedback loop (TTFL or pacemaker) with CLOCK as a conserved positive factor in animals. However, CLOCK’s functional evolutionary origin and mechanism of action in basal animals are unknown. In the cnidarianNematostella vectensis, pacemaker gene transcript levels, includingNvClk(theClockortholog), appear arrhythmic under constant darkness, questioning the role of NvCLK. Utilizing CRISPR/Cas9, we generated aNvClkallele mutant (NvClk^Δ), revealing circadian behavior loss in constant light conditions (LL and DD) while a 24-hour rhythm was maintained under light-dark condition (LD). Transcriptomics showed distinct rhythmic genes in wild-type (WT) genes in LD compared to DD. The LDNvClk^Δ/Δshowed comparable numbers of rhythmic genes, but were greatly reduced in DD. Furthermore, the LDNvClk^Δ/Δshowed alterations in temporal pacemaker gene expression, affecting their potential interactions. Additionally, differential expression of non-rhythmic genes associated with cell division and neuronal differentiation was observed. These findings revealed that a light-responsive pathway can partially compensate for circadian clock disruption and that theClockgene has evolved in cnidarians to synchronize rhythmic physiology and behavior to the diel rhythm of the earth’s biosphere.