Automated oligonucleotide solid-phase synthesis on nanosized silica particles using nano-on-micro assembled particle supports.
This article describes an original strategy to enable solid-phase oligodeoxyribonucleotide (ODN) synthesis on nanosized silica particles. It consists of the reversible immobilization of silica nanoparticles (NPs) on micrometric silica beads. The resulting assemblies, called nano-on-micro (NOM) systems, are well adapted to ODN synthesis in an automated instrument. First, NPs are derivatized with OH functions. For NOM assembly preparation, these functions react with the silanols of the microbeads under specific experimental conditions. Furthermore, OH groups allow ODN synthesis on the nanoparticles via phosphoramidite chemistry. The stability of the NOM assemblies during ODN solid-phase synthesis is confirmed by scanning and transmission electron microscopy (SEM and TEM, respectively), together with dynamic light scattering analyses. Then, the release of ODN-functionalized nanoparticles is performed under mild conditions (1% NH(4)OH in water, 1 h, 60 degrees C). Our technique provides silica nanoparticles well functionalized with oligonucleotides, as demonstrated by hybridization experiments conducted with the cDNA target.
Farre
Carole
C
Université de Lyon, Univ. Lyon 1, UMR 5180, Laboratoire des Sciences Analytiques, Villeurbanne, 69622 Cedex, France.
Lansalot
Muriel
M
Bazzi
Rana
R
Roux
Stéphane
S
Marquette
Christophe A
CA
Catanante
Gaëlle
G
Blum
Loïc J
LJ
Charvet
Nicolas
N
Louis
Cédric
C
Chaix
Carole
C
D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D008962:Models, Theoretical; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry
D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; D012822:Silicon Dioxide