By John Goodchild (auth.), John Goodchild (eds.)
The box of oligonucleotide therapeutics study is ripe with the chance of latest discoveries. In Therapeutic Oligonucleotides: equipment and Protocols, a variety of verified and rising tools for the applying of oligonucleotides as therapeutics are awarded, all offering the instruments had to encourage nice adjustments within the box. Divided into twenty-one chapters, this special quantity meticulously describes very important protocols for optimizing and bettering telephone uptake, reminiscent of photochemical internalization, changed cellphone penetrating peptides, antibody conjugates, and nanoparticles. different chapters tackle quantitation of RNA therapeutics in cells, assaying gene knockdown, selecting the right objective website and synthesis of assorted changed oligonucleotides. Written within the profitable Methods in Molecular Biology™ sequence layout, chapters contain introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, quite simply reproducible protocols, and notes on troubleshooting and keeping off recognized pitfalls.
Authoritative and simply available, Therapeutic Oligonucleotides: equipment and Protocols serves as a well timed source for either pros and rookies pursuing examine during this interesting and pioneering field.
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Extra info for Therapeutic Oligonucleotides: Methods and Protocols
And Cosstick, R. (1996) Synthesis and structure of S-nucleosidyl S-aryl disulfides and their reaction with phosphites. Tetrahedron 52, 1027–1034. , Fettes, K. , O’Neil, I. , and Cosstick, R. (2004) Synthesis of phosphorothioamidites derived from 3 -thio-3 -deoxythymidine and 3 -thio2 ,3 -dideoxycytidine and the automated synthesis of oligodeoxy-nucleotides containing a 3 -S-phosphorothiolate linkage. Nucleic Acids Res. 32, 495–501. Gaynor, J. , Piperakis, M. , and Cosstick, R. (2010) Reverse-direction (5 →3 ) synthesis of oligonucleotides containing a3 -S-phosphorothiolate linkage and 3 -terminal 3 -thionucleosides.
2. 4-N-Benzoyl-3 O-benzyl-5 -O-tertbutyldiphenylsily-5methyl-2 -O,4 -C(methyleneoxymethylene) cytidine, 11 1. 5 h. After addition of 28% aqueous ammonia solution, further stir the mixture at room temperature for 1 h. 2. After removal of the solvent under reduced pressure, extract the mixture with ethyl acetate. Wash the organic extracts with water and brine, dry over sodium sulfate, and concentrate under reduced pressure. 40 g, 87%). White powder; mp 56–58◦ C. 30 (2H, m). MS (FAB): m/z 732 (MH+ ).
72 (1H, brs). MS (FAB): m/z 692 (MH+ ). 7. 4-N-Benzoyl-3 O-[2-cyanoethoxy (diisopropylamino) phosphino]-5 -O-(4,4 dimethoxytrityl)-2 O,4 -C-(methyleneoxymethylene)cytidine, 22 1. 66 mmol) in anhydrous acetonitrile (20 mL) at room temperature and stir the mixture at room temperature for 20 h (see Note 6). 2. 43 mmol) and further stir the mixture for 6 h. 3. After addition of saturated aqueous sodium bicarbonate solution, extract the mixture with ethyl acetate. Wash the organic extracts with water and brine, dry over sodium sulfate, and concentrate under reduced pressure.
Therapeutic Oligonucleotides: Methods and Protocols by John Goodchild (auth.), John Goodchild (eds.)