title: DNA display III. Solid-phase organic synthesis on unprotected DNA.
abstract: DNA-directed synthesis represents a powerful new tool for molecular discovery. Its ultimate utility, however, hinges upon the diversity of chemical reactions that can be executed in the presence of unprotected DNA. We present a solid-phase reaction format that makes possible the use of standard organic reaction conditions and common reagents to facilitate chemical transformations on unprotected DNA supports. We demonstrate the feasibility of this strategy by comprehensively adapting solid-phase 9-fluorenylmethyoxycarbonyl-based peptide synthesis to be DNA-compatible, and we describe a set of tools for the adaptation of other chemistries. Efficient peptide coupling to DNA was observed for all 33 amino acids tested, and polypeptides as long as 12 amino acids were synthesized on DNA supports. Beyond the direct implications for synthesis of peptide-DNA conjugates, the methods described offer a general strategy for organic synthesis on unprotected DNA. Their employment can facilitate the generation of chemically diverse DNA-encoded molecular populations amenable to in vitro evolution and genetic manipulation.
authors.affiliation: Department of Biochemistry, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: David R
authors.initials: DR
authors.lastname: Halpin
authors.forename: Juanghae A
authors.initials: JA
authors.lastname: Lee
authors.forename: S Jarrett
authors.initials: SJ
authors.lastname: Wrenn
authors.forename: Pehr B
authors.initials: PB
authors.lastname: Harbury
mesh_terms: D002851:Chromatography, High Pressure Liquid; D020650:Combinatorial Chemistry Techniques; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D017931:DNA Primers / Q000737:chemistry; D004277:DNA, Single-Stranded / Q000737:chemistry; D004952:Esters / Q000737:chemistry; D005449:Fluorenes / Q000737:chemistry; D005818:Genetic Engineering; D005821:Genetic Techniques; D008956:Models, Chemical; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry; D015202:Protein Engineering; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000737:chemistry; D013388:Succinimides / Q000737:chemistry; D013696:Temperature
chemical_list: C496789:9-fluorenylmethoxycarbonyl; D017931:DNA Primers; D004277:DNA, Single-Stranded; D004952:Esters; D005449:Fluorenes; D009838:Oligodeoxyribonucleotides; D019151:Peptide Library; D010455:Peptides; D013388:Succinimides; D004247:DNA; D015719:Single-Strand Specific DNA and RNA Endonucleases