Research Update
NTPs Imparting Nuclease Resistance and More
2' Fluoro and 2' O-Methyl NTP analogs are being utilized in an increasing number of applications in research and new drug development. Recently 2' Fluoro dATP was confirmed to be a universal tool for screening ATP-requiring enzymes in the investigation of pharmaceutically applicable kinases.
However, 2' Fluoro and 2' O-Methyl modifications are most commonly incorporated for improvement of in vivo stability. They are used in enzymatic synthesis of aptamers, antagomirs and siRNA because they impart increased target affinity and nuclease resistance while reducing immune response. Furthermore, 2' O-Methyl modified sequences in particular form very stable chimeric duplexes with standard RNA.
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Analogs Currently Available
N-1007 2'-Fluoro-2'-dATP
N-1008 2'-Fluoro-2'-dCTP
N-1009 2'-Fluoro-2'-dGTP
N-1010 2'-Fluoro-2'-dUTP
N-1055 2’-Fluoro-2’-dTTP
N-1015 2'-O-Methyl-ATP
N-1016 2'-O-Methyl-CTP
N-1017 2'-O-Methyl-GTP
N-1018 2'-O-Methyl-UTP
N-1021 2'-O-Methyl-ITP
N-1040 2'-O-Methyl-2-amino-ATP
N-1041 2'-O-Methylpseudo-UTP
N-1043 2'-O-Methyl-5-methyl-UTP
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Technology Spotlight
Enhancing Oligonucleotide In Vivo Stability
Sulfurization of the phosphodiester backbone is the most frequently used method for enhancing in vivo stability of oligonucleotides. 2’OMe RNA substitution is another useful modification to create a more stable form of RNA with similar properties. It has the same low rate of depurination, but it is protected against base hydrolysis. Both phosphorothioate linkages and 2’OMe RNA backbones are commonly used in therapeutic applications. Other oligonucleotide modifications helpful in increasing in vivo nuclease resistance include 5’ Propyl, 2’,3’-Dideoxy modified bases, terminal phosphate groups and inverted bases. In a recent study by C. Roberts et al a 'roadblock' oligonucleotide, an oligo with the 3′ terminus modified with a chain-terminating propyl group, manufactured by TriLink was employed in short template amplicon and multiplex megaprimer-enabled relay (STAMMER) sequencing, a simultaneous approach to higher throughput sequence-based typing of polymorphic gene systems. The authors believe this method could significantly impact the output of sequence-based typing laboratories.
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Featured FAQ
Q - Which polymerases do you suggest for incorporation of 2’ OMe RNA and your other dNTPs/NTPs in transcription?
A - 2' Fluoro and 2' O-Methyl NTP analogs can be incorporated as substrates for RNA polymerases, with T7 R&DNA™ polymerase and SP6 R&DNA™ polymerases from Epicentre providing the best results. For many of the triphosphates we sell there is very little known. Please see our table of Enzymatic Activity of Selective NTPs, or our NTP bibliography to see if there is a publication regarding your application or analog of interest.
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