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Please use this identifier to cite or link to this item: http://hdl.handle.net/10805/1186

Title: “Spontaneous generation of RNA”
Authors: PINO, SAMANTA
Tutor: Di Mauro, Ernesto
Keywords: RNA
Ligation
abiotic polymerization
RNA stability
Issue Date: 12-Mar-2010
Abstract: The field of prebiotic chemistry deals with experimental and theoretical approaches and scenarios toward an understanding of how biologically relevant molecules may have been formed and how they may have developed into protocellular systems, which eventually led to the origin of life. In this context, many progresses has been made, in recent years, in the field of the origin of nucleobases, nucleotides and oligonucleotides. We have shown that formamide (HCONH2), one of the simplest molecules grouping the four most common elements of the universe H, C, O and N, provides a chemical frame potentially affording all the monomeric components necessary for the formation of nucleic polymers. In the presence of the appropriate catalysts, and by moderate heating, formamide yields a complete set of nucleic bases, acyclonucleosides and favours both phosphorylations and transphosphorylations. We also observed that nucleoside phosphorylation and oligomerization may take place in water in nonenzymatic abiotic conditions. Both observations contribute to the understanding of the origin of informational polymers. Finally, we present experiments showing the non enzymatic ligation of oligo RNAs in water leading to the formation of RNA of biologically relevant length, ready to undergo to Darwinian selection.
URI: http://hdl.handle.net/10805/1186
Research interests: The field of prebiotic chemistry deals with experimental and theoretical approaches and scenarios toward an understanding of how biologically relevant molecules may have been formed and how they may have developed into protocellular systems, which eventually led to the origin of life. In this context, many progresses has been made, in recent years, in the field of the origin of nucleobases, nucleotides and oligonucleotides. We have shown that formamide (HCONH2), one of the simplest molecules grouping the four most common elements of the universe H, C, O and N, provides a chemical frame potentially affording all the monomeric components necessary for the formation of nucleic polymers. In the presence of the appropriate catalysts, and by moderate heating, formamide yields a complete set of nucleic bases, acyclonucleosides and favours both phosphorylations and transphosphorylations. We also observed that nucleoside phosphorylation and oligomerization may take place in water in nonenzymatic abiotic conditions. Both observations contribute to the understanding of the origin of informational polymers. Finally, we present experiments showing the non enzymatic ligation of oligo RNAs in water leading to the formation of RNA of biologically relevant length, ready to undergo to Darwinian selection.
Skills short description: Cell cultures (human glioblastoma cells U-87MG, human bone tumor cells, human lymphoid cells, mouse fibroblasts) Cytofluorimetric assay of cell cycle and apoptosis by DNA labelling with Propidium Iodite (PI) Biparametric flow cytometric assay of cell cycle by incorporation of Bromodeoxyuridine (BrdU) Analysis of cell death (necrosis and apoptosis) of human lymphoid cells by using Trypan Blue Analysis of protein expression by acrylamide gel ( SDS-PAGE, Western blot) Extraction and purification of DNA from whole blood Amplification of DNA sequences by PCR Analysis and preparation of DNA fragments by agarose gel electrophoresis RFLP-PCR Comet assay on whole blood to evaluate DNA damages (SS and DS breaks) Evaluation of DNA damages by fluorescence microscopy CCD (Charged Couplet Device) High resolution nucleic acids electrophoresis Nucleic acids labelling with (32P) γATP Analysis of RNA interactions on mineral surface by Atomic Force Non-enzymatic generation of RNA from precursors
Appears in PhD:SCIENZE PASTEURIANE

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