Oligodeoxynucleotides protection

Noiri, E., Peresleni, T., Miller, F. and Goligorsky, M. S. (1996). In vivo targeting of inducible NO synthase with oligodeoxynucleotides protects rat kidney against ischemia. J. Clin. Invest. 97, 2377-2383. 

Poggio TV, La Torre JL, Scodeller EA (2006) Intranasal immunization with a recombinant truncated FimH adhesin adjuvanted with CpG oligodeoxynucleotides protects mice against uropathogenic Escherichia coli challenge. Can J Microbiol 52 1093-1102... 

Verthelyi DJ, Gursel M, Kenney RT et al (2(X)3) CpG oligodeoxynucleotides protect normal and SIV-infected macaques from Leishmania infection. J Immunol 170 4717-4723... 

Fig. 11. The amidite method for synthesizing oligodeoxynucleotides, where B and represent one of the protected bases thymine, A/ -benzoylcytosine,...

Previous studies have reported that ERKs are characteristically associated with cell proliferation and protection from apoptosis (Bl, XI), while activation of JNK and p38 MAPK can promote apoptosis in many systems, including B lymphocytes (G5), cerebellar granule cells (K3), hematopoietic cells (K8), and neuronal cells (M3, XI). On the other hand, a recent report found that a pyridinyl imidazole, SB 202190, the specific inhibitor of p38 MAPK, by itself was sufficient to induce apoptosis in T lymphocyte Jurkat cells (N2). Moreover, Th-2-derived cytokine IL-5, the ERK activator and antiapoptotic factor for eosinophils, could also activate p38 MAPK in human eosinophils (BIO). We recently reported that cytokine IL-3, IL-5, and GM-CSF could prolong survival of human eosinophilic leukemic (EoL-1) cells through the transient activation of ERK (W15). On the other hand, activation of p38 MAPK in EoL-1 cells by the NSAID sodium salicylate (NaSal) could lead to apoptosis (W15). We also found that the suppression of ERK using ERK antisense phosphorothioate oligodeoxynucleotides could promote the apoptosis of peripheral blood eosinophils (W16). Moreover, we found that dexamethasone-induced apoptosis and activation of JNK and p38 MAPK activity in eosinophils are regulated by caspases (Z2). 

Zimmermann, S., Egeter, O., Hausmann, S., Lipford, G.B., Rocken, M., Wagner, H. and Heeg, K. (1998) CpG oligodeoxynucleotides trigger protective and curative Thl responses in lethal murine leishmaniasis. J. Immunol., 160, 3627-3630. 

Dumais, N., et al. 2002. Mucosal immunization with inactivated human immunodeficiency virus plus CpG oligodeoxynucleotides induces genital immune responses and protection against intravaginal challenge. J Infect Dis 15 1098. 

Kwant, A., and K.L. Rosenthal. 2004. Intravaginal immunization with viral subunit protein plus CpG oligodeoxynucleotides induces protective immunity against HSV-2. Vaccine 22 3098. 

Chen H, Mohuczy D, Li D, et al. Protection against ischemia/reperfusion injury and myocardial dysfunction by anti-sense-oligodeoxynucleotide directed at angiotensin-converting enzyme mRNA. Gene Ther 2001 8 804-810. 

There are three widely used chemistries for oligodeoxynucleotide synthesis, and all have been successfully transferred onto SP. The first is the so-called phosphotriester approach (71), which is shown in Fig. 2.9. In this method a 5 -protected nucleoside... 

The synthesis and incorporation into oligonucleotides of the A-phosphory-lated deoxycytidine 3 -phosphoramidites (144) and (145) obtained either from the O-protected 2 -deoxycytidine and bis(2-cyanoethyl) A,A-diisopropyl phosphoramidite or diethylphosphorochloridite, respectively, was described by Sekine. Sekine also reported that the A-phosphorylated derivatives of 2 -deoxy adenosine decomposed readily and were unsuitable for incorporation into oligodeoxynucleotides. 

The synthesis and hybridisation properties of oligodeoxynucleotides containing 7-( 1 -propynyl)-7-deaza-2 -deoxyguanosine and 7-( 1 -propynyl)-7-deaza-2 -deoxya-denosine have been described. The suitably protected nucleosides 199a and 199b were synthesised and incorporated into oligomers. Thermal denaturation of these oligomers hybridised to RNA demonstrates an increased stability relative to 7-unsubstituted deazapurine and unmodified controls. 

In order to preserve optimal stability in the hybridization of RNA by sugar-modified oligonucleotides, nucleosides substituted at the 2 -position must retain a C3 -endo puckered conformation. To this end, the 2 -0- 2-[iV,N-(dimethylamino)oxy]ethyl and the 2 -0- 2-[Ar,lV-(diethylamino)oxy]-ethyl 3 -phosphoramidite derivatives of thymidine (38a, 38b) have been synthesised. The nucleoside precursors were prepared from a phthalimido-derivative obtained from 2,2 -anhydro-5-methyluridine which had been treated, after appropriate protection, with a borate ester generated in situ and then reacted with PPha, DEAD and AT-hydroxyphthalimide. The incorporation of (38a) and (38b) in oligonucleotides and oligodeoxynucleotides resulted in high binding affinity for... 

It has been demonstrated that it is possible to insert paramagnetic probes into nucleic acids in a site- and type-specific manner. To this end, the 2,2,6,6-tet-ramethyl-l-piperidinyloxy free radical-labelled phosphoramidites of 2-amino-2 -deoxyadenosine, 2 -deoxyadenosine, 2 -deoxycytosine and 5-methyl-2 -deoxycytosine have been prepared and used for the automatic synthesis of mono-labeled oligodeoxynucleotides which proved active by EPR. The phosphoramidites carry a nitroxide spin-label directly linked to the exocyclic amino group. Starting from appropriately protected 2 -deoxyguanosine or 2 -deoxyinosine, the precursors to (66a) and (66b) were obtained by nucleophilic... 

Phosphorus protection. The protecting group is stable to acid and is removed by fragmentation using aqueous NH4OH. It is introduced by the phosphoramidite approach and is useful for internucleotidic bonds in oligodeoxynucleotide synthesis. 

The deprotection of crude oligodeoxynucleotides traditionally requires a basic treatment that allows the concomitant removal of the exocyclic acyl protecting groups, -elimination of the 2-cyanoethyl phosphate protecting group, and cleavage of the succinic ester bond that links the oligonucleotide to the solid support (Fig. 9). 

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