1. Nonenzymatic RNA-templated Synthesis of N3'→P5' Phosphoramidate DNA.
- Author
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O'Flaherty DK, Zhou L, and Szostak JW
- Abstract
The RNA world hypothesis describes a scenario where early life forms relied on RNA to govern both inheritance and catalyze useful chemical reactions. Prior to the emergence of enzymes capable of replicating the RNA genome, a nonenzymatic replication process would have been necessary to initiate Darwinian Evolution. However, the one-pot nonenzymatic RNA chemical copying of templates with mixed-sequences is insufficient to generate strand products long enough to encode useful function. The use of alternate (RNA-like) genetic polymers may overcome hurdles associated with RNA copying, and further our understanding of nonenzymatic copying chemistry. This protocol describes the nonenzymatic copying of RNA templates into N3'→P5' phosphoramidate DNA (3'-NP-DNA). We describe, in detail, the synthesis of 3'-amino-2',3'-dideoxyribonucleotide monomers activated with 2-aminoimidazole (3'-NH
2 -2AIpddN), and their use in template-directed polymerization., Competing Interests: Competing interestsThe authors declare no competing financial interest., (Copyright © 2020 The Authors; exclusive licensee Bio-protocol LLC.)- Published
- 2020
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