TY - JOUR
T1 - An in vitro translation, selection and amplification system for peptide nucleic acids
AU - Brudno, Yevgeny
AU - Birnbaum, Michael E.
AU - Kleiner, Ralph E.
AU - Liu, David R.
N1 - Funding Information:
This research was supported by the US Office of Naval Research (N00014-03-1-0749), the National Institutes of Health (R01GM065865) and the Howard Hughes Medical Institute. Y.B. gratefully acknowledges the support of an NSF Graduate Research Fellowship.
PY - 2010/2
Y1 - 2010/2
N2 - Methods to evolve synthetic, rather than biological, polymers could significantly expand the functional potential of polymers that emerge from in vitro evolution. Requirements for synthetic polymer evolution include (i) sequence-specific polymerization of synthetic building blocks on an amplifiable template, (ii) display of the newly translated polymer strand in a manner that allows it to adopt folded structures, (iii) selection of synthetic polymer libraries for desired binding or catalytic properties and (iv) amplification of template sequences that survive selection in a manner that allows subsequent translation. Here we report the development of such a system for peptide nucleic acids (PNAs) using a set of 12 PNA pentamer building blocks. We validated the system by performing six iterated cycles of translation, selection and amplification on a library of 4.3 ×- 108 PNA-encoding DNA templates and observed > 1,000,000-fold overall enrichment of a template encoding a biotinylated (streptavidin-binding) PNA. These results collectively provide an experimental foundation for PNA evolution in the aboratory.
AB - Methods to evolve synthetic, rather than biological, polymers could significantly expand the functional potential of polymers that emerge from in vitro evolution. Requirements for synthetic polymer evolution include (i) sequence-specific polymerization of synthetic building blocks on an amplifiable template, (ii) display of the newly translated polymer strand in a manner that allows it to adopt folded structures, (iii) selection of synthetic polymer libraries for desired binding or catalytic properties and (iv) amplification of template sequences that survive selection in a manner that allows subsequent translation. Here we report the development of such a system for peptide nucleic acids (PNAs) using a set of 12 PNA pentamer building blocks. We validated the system by performing six iterated cycles of translation, selection and amplification on a library of 4.3 ×- 108 PNA-encoding DNA templates and observed > 1,000,000-fold overall enrichment of a template encoding a biotinylated (streptavidin-binding) PNA. These results collectively provide an experimental foundation for PNA evolution in the aboratory.
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U2 - 10.1038/nchembio.280
DO - 10.1038/nchembio.280
M3 - Article
C2 - 20081830
AN - SCOPUS:75349085231
SN - 1552-4450
VL - 6
SP - 148
EP - 155
JO - Nature Chemical Biology
JF - Nature Chemical Biology
IS - 2
ER -