Pyrophosphate-Mediated Repair of Damaged and Mismatched RNA by a Polymerase Ribozyme

Prior to the emergence of the contemporary biosphere, the first replicating systems are thought to have progressed through an RNA-based stage. Such an evolving world would likely have transferred heritable information during replication using RNA polymerase ribozymes. Though substantial effort has been put forth towards evolving RNA polymerases, many variants suffer from premature termination and low fidelity, resulting in low yields of full-length or active sequences. Replication of longer sequences requires a sufficiently high fidelity to lend an evolutionary advantage to an evolvable system. Here we demonstrate ribozyme-mediated repair of mismatched and damaged RNA sequences. Under conditions of saturating pyrophosphate concentrations, we show that a polymerase ribozyme can repair RNA sequences terminated in a mismatch, a non-extendable 2'-3' cyclic phosphate, or both, to generate a triphosphorylated nucleotide. This repair step increases the fidelity and allows polymerization along an extended template, including the ribozyme itself. This increase of copying fidelity advances the longstanding goal of developing a self-replicating polymerase ribozyme.