title: Chemical Morphing of DNA Containing Four Noncanonical Bases.
abstract: The ability of alternative nucleic acids, in which all four nucleobases are substituted, to replicate in vitro and to serve as genetic templates in vivo was evaluated. A nucleotide triphosphate set of 5-chloro-2'-deoxyuridine, 7-deaza-2'-deoxyadenosine, 5-fluoro-2'-deoxycytidine, and 7-deaza-2'deoxyguanosine successfully underwent polymerase chain reaction (PCR) amplification using templates of different lengths (57 or 525mer) and Taq or Vent (exo-) DNA polymerases as catalysts. Furthermore, a fully morphed gene encoding a dihydrofolate reductase was generated by PCR using these fully substituted nucleotides and was shown to transform and confer trimethoprim resistance to E. coli. These results demonstrated that fully modified templates were accurately read by the bacterial replication machinery and provide the first example of a long fully modified DNA molecule being functional in vivo.
authors.affiliation: Laboratory of Medicinal Chemistry, Rega, Institute for Medical Research, KU Leuven, Minderbroedersstraat 10, 3000, Leuven, Belgium.
authors.forename: Elena
authors.initials: E
authors.lastname: Eremeeva
authors.affiliation: Laboratory of Medicinal Chemistry, Rega, Institute for Medical Research, KU Leuven, Minderbroedersstraat 10, 3000, Leuven, Belgium.
authors.forename: Michail
authors.initials: M
authors.lastname: Abramov
authors.affiliation: Laboratory of Medicinal Chemistry, Rega, Institute for Medical Research, KU Leuven, Minderbroedersstraat 10, 3000, Leuven, Belgium.
authors.forename: Lia
authors.initials: L
authors.lastname: Margamuljana
authors.affiliation: Laboratory of Medicinal Chemistry, Rega, Institute for Medical Research, KU Leuven, Minderbroedersstraat 10, 3000, Leuven, Belgium.
authors.forename: Jef
authors.initials: J
authors.lastname: Rozenski
authors.affiliation: ISSB, Génopole, Genavenir 6, Equipe Xénome, 5 rue Henri Desbruères, 91030, Evry Cedex, France.
authors.forename: Valerie
authors.initials: V
authors.lastname: Pezo
authors.affiliation: ISSB, Génopole, Genavenir 6, Equipe Xénome, 5 rue Henri Desbruères, 91030, Evry Cedex, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Marlière
authors.affiliation: Laboratory of Medicinal Chemistry, Rega, Institute for Medical Research, KU Leuven, Minderbroedersstraat 10, 3000, Leuven, Belgium. Piet.Herdewijn@rega.kuleuven.be.|ISSB, Génopole, Genavenir 6, Equipe Xénome, 5 rue Henri Desbruères, 91030, Evry Cedex, France. Piet.Herdewijn@rega.kuleuven.be.
authors.forename: Piet
authors.initials: P
authors.lastname: Herdewijn
mesh_terms: D004247:DNA / Q000737:chemistry*; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000737:chemistry; D003848:Deoxyguanine Nucleotides / Q000737:chemistry; D003857:Deoxyuridine / Q000031:analogs & derivatives / Q000737:chemistry; D004926:Escherichia coli / Q000187:drug effects; D016133:Polymerase Chain Reaction / Q000379:methods; D014295:Trimethoprim / Q000633:toxicity; D014296:Trimethoprim Resistance; D014372:Tubercidin / Q000031:analogs & derivatives / Q000737:chemistry
chemical_list: D003848:Deoxyguanine Nucleotides; D003841:Deoxycytidine; C065361:2'-deoxy-7-deazaguanosine triphosphate; C012244:5-chloro-2'-deoxyuridine; C053222:2'-deoxytubercidin; D004247:DNA; D014295:Trimethoprim; C007746:5-fluoro-2'-deoxycytidine; D014372:Tubercidin; D003857:Deoxyuridine
keywords: DNA replication; chemical evolution; gene expression; nucleic acids; synthetic biology