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Circular mRNA can direct translation of extremely long repeating-sequence proteins in vivo

Summary: Circular mRNA can direct translation of extremely
long repeating-sequence proteins in vivo
Center for Molecular Biology of RNA, Biology Department, Sinsheimer Laboratories, University of California, Santa Cruz,
Santa Cruz, California 95064, USA
Many proteins with unusual structural properties are comprised of multiple repeating amino acid sequences and are
often fractious to expression in recombinant systems. To facilitate recombinant production of such proteins for
structural and engineering studies, we have produced circular messenger RNAs with infinite open reading frames. We
show that a circular mRNA containing a simple green fluorescent protein (GFP) open reading frame can direct GFP
expression in Escherichia coli. A circular mRNA with an infinite GFP open reading frame produces extremely long
protein chains, proving that bacterial ribosomes can internally initiate and repeatedly transit a circular mRNA. Only
the monomeric forms of GFP produced from circular mRNA are fluorescent. Analysis of the translation initiation
region shows that multiple sequences contribute to maximal translation from circular mRNA. This technology pro-
vides a unique means of producing a very long repeating-sequence protein, and may open the way for development
of proteinaceous materials with novel properties.
Keywords: circular mRNA; long repeating protein; translation
All known natural eukaryotic and prokaryotic messen-
ger RNAs are linear+ Although circular RNAs exist in


Source: Ares Jr., Manny - Department of Molecular, Cell, and Developmental Biology, University of California at Santa Cruz


Collections: Biology and Medicine