In his March 7, 2017 Zone-in-with-Zon blog post, “Evolving Polymerases to Do the Impossible” (http://zon.trilinkbiotech.com/), Dr. Jerry Zon describes recent work (http://www.nature.com/nchem/journal/v8/n6/full/nchem.2493.html) of Professor Floyd Romesberg of the Scripps Research Institute Department of Chemistry, and his team, as “a tour de force of experimental methodology” that has evolved nucleic acid polymerases into mutant polymerases that can do what previously seemed impossible. Romesberg’s lab has, Dr. Zon writes, “successfully achieved what I think of as ‘multiple molecular moonshots,’ wherein a Taq polymerase (which normally reads and writes DNA during PCR), was evolved by novel selection (SELEX) methods into mutant polymerases that are able to transcribe DNA into 2’-O-methyl (2’-OMe) RNA, and reverse-transcribe 2’-OMe RNA into DNA for PCR/sequencing…This was exemplified using a 60-mer DNA template and 18-mer 2’-OMe RNA primer to produce a fully-modified 48-mer 2’-OMe RNA by means of an evolved mPol and all four A, G, C and U 2’-OMe NTPs, which I am proud to say were purchased from TriLink BioTechnologies! This type of molecular evolution of a polymerase has no precedent.” With regard to the significance of this achievement, Dr. Zon said the following. “The present study is the first to evolve an mPol for reverse transcription of fully modified 2’-OMe RNA into DNA, which can then be amplified by PCR and/or sequenced, thereby opening the door for a variety of new analytical methods. Most importantly, the molecular mechanism by which these remarkable mPol activities were evolved, namely, the stabilization of an interaction between the ‘thumb and fingers domains,’ may be general and thus useful for the optimization of other Pols.
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