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Significance We describe the smallest (220 nt) replicating covalently closed circular (CCC) RNA (nanogenome) that contains numerous unique and novel properties of coding and translation. It is the smallest of all viroids and virusoids and the only one to encode proteins. No noncoding sequences could be identified. Initiation and termination codons are combined in UGAUGA, and read-throughs generate larger proteins. This is the only CCC RNA to be directly translated by eukaryotic ribosomes. It also contains an internal ribosome entry site and it defies the classical Kozak scanning model for initiation of translation. Is it a living fossil or is it a novel trend in genomic evolution?

Novel coding, translation, and gene expression of a replicating covalently closed circular RNA of 220 nt