The Joyce laboratory uses directed evolution to develop RNA enzymes that replicate RNA, aiming to construct RNA-based life in the laboratory and enabling the experimental exploration of the RNA world that is thought to have preceded DNA and protein-based cellular organisms during the early history of life on Earth. Toward this goal, a highly active polymerase ribozyme, termed ‘24-3’ was evolved that can synthesize RNA from RNA templates, but was also found to be capable of synthesizing DNA from RNA templates. The unselected emergence of this reverse transcriptase activity suggests that the transition from an RNA world to one using DNA as a more stable repository of genetic information could have preceded the emergence of genetically encoded proteins. Multiple rounds of directed evolution have been carried out with the aim of improving this modest reverse transcriptase activity. Iterative sequence-selective restriction enzyme digestion of ribozyme-extended DNA has furnished reverse transcriptase ribozymes with enhanced activity.<br/>

Making DNA from RNA: Optimizing the reverse transcriptase activity of an evolved ribozyme

2019 Fall National Meeting

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