Faster and cleaner dynamic kinetic resolution via mechanochemistry

International audienceApplication of the ball-milling techniques to dynamic kinetic resolution accelerates reactions while avoiding the use of toxic organic solvents and reactants commonly required in these processes. In this way, dynamic kinetic resolutions can be both faster and “cleaner” in the sense that mechanochemistry enables the reduction of their environmental impact

Study of the biocompatibility of nucleic acid modified by boronic acid : development to new diagnostic tools

La modification d’oligonucléotides est un domaine attrayant de la chimie organique. De nombreuses études se sont intéressées à la génération de liens internucléosidiques artificiels à visée thérapeutique, diagnostic ou encore pour des applications en chimie prébiotique. Ce manuscript de thèse rapporte la synthèse et l’étude de biocompatibilité d’acides nucléiques modifiés à leurs extrémités 5’ par un acide boronique. Les comportement des oligomères boroniques a été évalué en présence de différentes classes d’enzymes telles que les ligases, les polymérases ou encore les phosphodiestérases. Les résultats de biocompatibilité obtenus en présence de ces enzymes nous ont permis d’utiliser ces acides nucléiques modifiés comme de réels outils de diagnostic pour réaliser de la détection de point de mutation ou encore de la détection de péroxynitrite in-cellulo.The modification of oligonucleotides is an attractive field of organic chemistry. Many studies have focused on the generation of artificial internucleoside linkages for therapeutic, diagnostic or for applications in prebiotic chemistry. This thesis manuscript reports the synthesis and study of nucleic acids biocompatibility modified at their 5 'ends by a boronic acid function. The behavior of boronic oligomers was assessed in the presence of different classes of enzymes, such as ligases, polymerases or phosphodiesterases. The biocompatibility results obtained in the presence of these enzymes allowed us to use these modified nucleic acids as real diagnostic tools to achieve mutation point detection or detection of peroxynitrite in-cellulo

Nuclease stability of boron-modified nucleic acids: application to label-free mismatch detection

International audience5′-End boronic acid-modified oligonucleotides were evaluated against various nucleases at single and double stranded levels. The results show that these modifications induce a high resistance to degradation by calf-spleen and snake venom phosphodiesterases. More importantly, this eventually led to the development of a new label-free enzyme-assisted fluorescence-based method for single mismatch detection

RNA-directed off/on switch of RNase H activity using boronic ester formation

International audienceRNase H is a non-specific endonuclease which degrades selectively the RNA strand in DNA/RNA duplexes. We demonstrate in the present study that 5'-boronic acid modified oligonucleotides hybridized to a RNA target sequence converts RNase H to an inactivated enzyme complex. The dynamic formation of a boronate ester upon addition of a diol moiety disrupts the enzyme-inhibitor complex and reactivates RNase H. Moreover, we show that reactivation of RNase H function can also be engineered through short RNA trimers inputs that fashion RNase H from a non-specific DNA-guided enzyme into an informational and programmable RNA-guided one. Examples of programmable RNA recognition and cleavage illustrate the potential of this new stimuli-responsive system