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An in vitro translation, selection and amplification system for peptide nucleic acids
Yevgeny Brudno1, Michael E Birnbaum, Ralph E Kleiner
1Department of Chemistry and Chemical Biology, Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.
Nature Chemical Biology
|January 19, 2010
Summary
Researchers developed a novel system for evolving synthetic polymers, specifically peptide nucleic acids (PNAs). This breakthrough enables the in vitro evolution of PNAs, expanding their functional potential for various applications.
Area of Science:
- Synthetic polymer chemistry
- Molecular biology
- Biotechnology
Background:
- In vitro evolution typically uses biological polymers, limiting functional scope.
- Synthetic polymer evolution requires sequence-specific polymerization, display, selection, and amplification.
- Peptide nucleic acids (PNAs) offer unique chemical properties for synthetic polymer applications.
Purpose of the Study:
- To develop a system for the in vitro evolution of synthetic polymers, specifically PNAs.
- To enable sequence-specific polymerization and selection of PNAs based on desired properties.
- To establish a foundation for expanding the functional repertoire of synthetic polymers.
Main Methods:
- Developed a system for PNA evolution using 12 PNA pentamer building blocks.
- Utilized an amplifiable DNA template for sequence-specific PNA polymerization.
- Integrated translation, selection, and amplification of PNA-encoding DNA templates.
- Performed six iterated cycles of evolution on a library of 4.3 x 10^8 PNA-encoding DNA templates.
Main Results:
- Successfully demonstrated in vitro evolution of peptide nucleic acids (PNAs).
- Achieved over 1,000,000-fold enrichment of a PNA template encoding a biotinylated PNA.
- Validated the system's capability for iterative selection and amplification of functional PNAs.
- Showcased the potential for PNA libraries to be selected for specific binding properties.
Conclusions:
- Established an experimental foundation for the laboratory evolution of PNAs.
- The developed system significantly expands the potential for synthetic polymer functionalization.
- This work paves the way for novel applications of engineered synthetic polymers.
