NUCLEASE RESISTANT RIBOZYMES WITH HIGH CATALYTIC ACTIVITY

被引:144
作者
PAOLELLA, G
SPROAT, BS
LAMOND, AI
机构
[1] EMBL, D-6900 Heidelberg
关键词
HAMMERHEAD; RIBOZYMES; RNA CATALYSIS; RIBONUCLEOTIDE ANALOGS; 2'-O-ALKYLRIBONUCLEOTIDES;
D O I
10.1002/j.1460-2075.1992.tb05244.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Hammerhead ribozymes are efficient RNA enzymes characterized by a typical hammerhead secondary structure and a number of conserved bases. Little is known about the role of the ribose-phosphate backbone, although it is obviously important since a DNA molecule with the same base sequence is not a catalyst. Here we describe the synthesis of artificial ribozymes where modified (2'-O-allyl- and 2'-O-methyl-) ribonucleotides substitute for the corresponding ribonucleotides. A systematic analysis of partially substituted polymers identified a minimum set of six non-contiguous positions where insertion of modified ribonucleotides strongly affects catalytic activity. Surprisingly, ribozymes completely substituted except for these six ribonucleotides are still very active. These molecules efficiently cleave in trans target RNAs in a sequence-specific way, but, unlike RNA ribozymes, are very resistant to nuclease degradation and are very stable in serum. These properties make such synthetic polymers potentially useful for in vivo gene expression studies and therapeutic applications.
引用
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页码:1913 / 1919
页数:7
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