title: DNA ligase-mediated translation of DNA into densely functionalized nucleic acid polymers.
abstract: We developed a method to translate DNA sequences into densely functionalized nucleic acids by using T4 DNA ligase to mediate the DNA-templated polymerization of 5'-phosphorylated trinucleotides containing a wide variety of appended functional groups. This polymerization proceeds sequence specifically along a DNA template and can generate polymers of at least 50 building blocks (150 nucleotides) in length with remarkable efficiency. The resulting single-stranded highly modified nucleic acid is a suitable template for primer extension using deep vent (exo-) DNA polymerase, thereby enabling the regeneration of template DNA. We integrated these capabilities to perform iterated cycles of in vitro translation, selection, and template regeneration on libraries of modified nucleic acid polymers.
authors.affiliation: Howard Hughes Medical Institute, Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts, 02138, United States.
authors.forename: Ryan
authors.initials: R
authors.lastname: Hili
authors.forename: Jia
authors.initials: J
authors.lastname: Niu
authors.forename: David R
authors.initials: DR
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D017122:Bacteriophage T4 / Q000201:enzymology; D055162:Biocatalysis; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D011088:DNA Ligases / Q000737:chemistry / Q000378:metabolism*; D009696:Nucleic Acids / Q000737:chemistry / Q000378:metabolism*; D011108:Polymers / Q000737:chemistry / Q000378:metabolism*
chemical_list: D009696:Nucleic Acids; D011108:Polymers; D004247:DNA; D011088:DNA Ligases