Discovery, Characterization, and Optimization of an Unnatural Base Pair for Expansion of the Genetic Alphabet

DNA is inherently limited by its four natural nucleotides. Efforts to expand the genetic alphabet, by addition of an unnatural base pair, promise to expand the biotechnological applications available for DNA as well as to be an essential first step toward expansion of the genetic code. We have condu...

Full description

Saved in:
Bibliographic Details
Published in:Journal of the American Chemical Society 2008-02, Vol.130 (7), p.2336-2343
Main Authors: Leconte, Aaron M, Hwang, Gil Tae, Matsuda, Shigeo, Capek, Petr, Hari, Yoshiyuki, Romesberg, Floyd E
Format: Article
Language:English
Subjects:
Base Pairing
Hydrophobic and Hydrophilic Interactions
Nucleotides - chemical synthesis
Nucleotides - chemistry
Nucleotides - genetics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:DNA is inherently limited by its four natural nucleotides. Efforts to expand the genetic alphabet, by addition of an unnatural base pair, promise to expand the biotechnological applications available for DNA as well as to be an essential first step toward expansion of the genetic code. We have conducted two independent screens of hydrophobic unnatural nucleotides to identify novel candidate base pairs that are well recognized by a natural DNA polymerase. From a pool of 3600 candidate base pairs, both screens identified the same base pair, dSICS:dMMO2, which we report here. Using a series of related analogues, we performed a detailed structure−activity relationship analysis, which allowed us to identify the essential functional groups on each nucleobase. From the results of these studies, we designed an optimized base pair, d5SICS:dMMO2, which is efficiently and selectively synthesized by Kf within the context of natural DNA.
ISSN:0002-7863
1520-5126
DOI:10.1021/ja078223d