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Oligo Synthesis

Oligo Synthesis : CEPs

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2'-F-Ac-C-ANA-CE Phosphoramidite

2'-F-Ac-C-ANA-CE Phosphoramidite 5'-Dimethoxytrityl-N4-acetyl-2'-deoxy-2'-fluoroarabinocytidine, 3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite

Glen Research

Catalogue No.DescriptionPack SizePriceQty
10-3815-022'-F-Ac-C-ANA-CE Phosphoramidite0.25 g £482.00 Quantity Add to Order
10-3815-052'-F-Ac-C-ANA-CE Phosphoramidite0.5 g £955.00 Quantity Add to Order

Description

Arabinonucleosides are epimers of ribonucleosides with the chiral switch being at the 2' position of the sugar residue.   2'-F-ANA adopts a more DNA-like β-type helix conformation, not through the typical C2'-endo conformation but, rather, through an unusual O4'-endo (east) pucker.  However, the presence of the electronegative fluorine leads to a still significant increase (DTm1.2°C/mod) in melting temperature per modification.1 2'-F-ANA-containing oligonucleotides exhibit very high binding specificity to their targets.  Indeed, a single mismatch in a 2'-F-ANA - RNA duplex leads to a DTm of -7.2°C and in a 2'-F-ANA - DNA duplex a DTm of -3.9°C.2  

The presence of fluorine at the 2' position in 2'-F-ANA leads to increased stability to hydrolysis under basic conditions relative to RNA and even 2'-F-RNA.1,3   The stability of 2'-F-ANA to nucleases also makes this a useful modification for enhancing the stability of oligonucleotides in biological environments.2  2'-F-ANA hybridizes strongly to target RNA and, unlike most 2' modifications, induces cleavage of the target by RNase H.  Phosphorothioate (PS) 2'-F-ANA is routinely used in these applications due to its increased nuclease resistance.  Alternating 2'-F-ANA and DNA units provide among the highest potency RNase H-activating oligomers.  Both the "altimer" and "gapmer" strand architectures consistently outperform PS-DNA and DNA/RNA gapmers.4 

siRNA oligos were found to tolerate the presence of 2'-F-ANA linkages very well.  High potency gene silencing was demonstrated5 with siRNA chimeras containing 2'-F-RNA and/or LNA and 2'-F-ANA.  The high efficacy of these chimeras was attributed to the combination of the rigid RNA-like properties of 2'-F-RNA and LNA with the DNA-like properties of 2'-F-ANA.
 
 
Formula: C41H49FN5O8
M.W.: 789.83 
F.W.: 307.17 

Diluent: Anhydrous Acetonitrile

Coupling: 6 minute coupling time recommended.

Deprotection: No changes needed from standard method recommended by synthesizer manufacturer.

Storage: Storage: Refrigerated storage, maximum of 2-8°C, dry

Stability in Solution: 1-2 days

Please Note: 2'-F-ANA is covered by intellectual property.  Key patents covering siRNA and antisense applications are as follows: WO/2009/146556 (siRNA); WO 03064441 and WO 0220773 (antisense).

 

  1. E . Viazovkina, M.M. Mangos, M.I. Elzagheid, and M.J. Damha, Curr Protoc Nucleic Acid Chem, 2002, Chapter 4, Unit 4 15.
  2. J.K. Watts, and M.J. Damha, Can. J. Chem., 2008, 86, 641-656.
  3. J.K. Watts, A. Katolik, J. Viladoms, and M.J. Damha, Org Biomol Chem, 2009, 7, 1904-10.
  4. A. Kalota, et al., Nucleic Acids Res., 2006, 34, 451.
  5. G.F. Deleavey, et al., Nucleic Acids Res., 2010, 38, 4547-4557, J.K. Watts, et al., Nucleic Acids Res., 2007, 35, 1441-1451, T. Dowler, et al., Nucleic Acids Res., 2006, 34, 1669-1675.

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Protocols

MSDS

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References

  • E . Viazovkina, M.M. Mangos, M.I. Elzagheid, and M.J. Damha, Curr Protoc Nucleic Acid Chem, 2002, Chapter 4, Unit 4 15.
  • J.K. Watts, and M.J. Damha, Can. J. Chem., 2008, 86, 641-656.
  • J.K. Watts, A. Katolik, J. Viladoms, and M.J. Damha, Org Biomol Chem, 2009, 7, 1904-10.
  • A. Kalota, et al., Nucleic Acids Res., 2006, 34, 451.
  • G.F. Deleavey, et al., Nucleic Acids Res., 2010, 38, 4547-4557, J.K. Watts, et al., Nucleic Acids Res., 2007, 35, 1441-1451, T. Dowler, et al., Nucleic Acids Res., 2006, 34, 1669-1675.

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Notes

Stability note

Synthetic oligonucleotides containing 2'-F-RNA linkages may be deprotected with ammonium hydroxide as normal. Deprotection using AMA at 65°C leads to some degradation and so we recommend the use of AMA at room temperature for 2 hours.

 

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Applications & Benefits

DILUTION/COUPLING DATA

The table below shows pack size data and, for solutions, dilutions and approximate couplings based on normal priming procedures. Please link for more detailed usage information with the various synthesizers.

ABI 392/394
Cat.No. Pack Size Grams/ Pack 0.1M Dil. (mL) LV40 LV200 40nm 0.2µm 1µm 10µm
Approximate Number of Additions
10-3815-02 0.25grams .25grams 3.17 92.33 55.4 34.63 25.18 18.47 4.62
10-3815-05 0.5grams .5grams 6.33 197.67 118.6 74.13 53.91 39.53 9.88
Expedite
Cat.No. Pack Size Grams/ Pack Dilution (mL) Molarity 50nm 0.2µm 1µm 15µm  
Approximate Number of Additions
10-3815-02 0.25grams .25grams 4.72 .07 88 55 40 5.5  
10-3815-05 0.5grams .5grams 9.45 .07 182.6 114.13 83 11.41  
Mermade
Cat.No. Pack Size Grams/ Pack Dilution (mL) Molarity 30nm 200nm 1000nm    
Approximate Number of Additions
10-3815-02 0.25grams .25grams 4.72 .07 89.6 56 40.73    
10-3815-05 0.5grams .5grams 9.45 .07 184.2 115.13 83.73    

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Related products

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