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Activity and Stability of Hammerhead Ribozymes Containing 2′-C-Methyluridine: a New RNA Mimic

✍ Scribed by Mariana Gallo; Rosel Kretschmer-Kazemi Far; Georg Sczakiel; Adolfo M. Iribarren


Publisher
John Wiley and Sons
Year
2005
Tongue
English
Weight
72 KB
Volume
2
Category
Article
ISSN
1612-1872

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✦ Synopsis


We propose 2'-C-methylnucleotides as a new class of 2'-modified RNA mimics. These analogues are expected to provide 2'-OH groups capable of reproducing the interactions observed in natural RNA and, due to the presence of the Me group, to possess increased stability towards nucleases. In this work, we investigate the catalytic activity and nuclease resistance of hammerhead ribozymes carrying 2'-C-methyluridines in positions 4 and 7 of the catalytic core. We describe the in vitro activity of these chimeric molecules and their stability in cell lysate, fetal calf serum, and cell culture medium. The data show that, when only position 4 is modified, activity decreases twofold; while, when both 4 and 7 positions are substituted, a sevenfold drop in activity is observed. Regarding biological stability, the main increase of the half-life time is observed when position 7 is modified. These results suggest that 2'-C-methylnucleotides may be useful in the design of chemically synthesized RNA mimics with biological activity.