Postsynthetic modifications of DNA and RNA by bioorthogonal reactions

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In the first funding period, the copper-catalzed azide-acetylen cycloaddition, the strain-promoted azide-acetylen cycloaddtion, the photoclick reaction and the inverse electron demand Diels-Alder reaction were implemented as important bioorthogonal modifications into deoxyuridines as phosphoramidites for the chemical DNA synthesis and as triphosphates for the enzymatic DNA preparation. Using the strain-promoted cycloaddition, the successfull postsynthetic fluorescent labeling inside living cells was demonstrated as a proof-of-principle. Based on these substantial results the bioorthogonally reactive groups are further developed for postsynthetic modifications of DNA and RNA and applied inside living cells. More precisely, the second funding period includes the usage of visible light for the photoclick reactions, the development of rather small reactive groups, the application in PCR, the modification of RNA and the evulation and application of these reactions in living cells.