Kinetic selectivity of complementary nucleic acids: bcr-abl-directed antisense RNA and ribozymes
Efficacy and sequence specificity are two major requirements in the use of antisense nucleic acids and ribozymes. For long-chain complementary RNA sequences (>30 nt), effects in living cells are correlated with the association rate of the complementary RNAin vitro, but not with the stability of t...
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| Main Authors: | , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
21 June 1996
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| In: |
Journal of molecular biology
Year: 1996, Volume: 259, Issue: 4, Pages: 632-644 |
| ISSN: | 1089-8638 |
| DOI: | 10.1006/jmbi.1996.0345 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1006/jmbi.1996.0345 Verlag, lizenzpflichtig, Volltext: https://www.sciencedirect.com/science/article/pii/S0022283696903459 
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| Author Notes: | Ralf Kronenwett, Rainer Haas, Georg Sczakiel |
MARC
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| 245 | 1 | 0 | |a Kinetic selectivity of complementary nucleic acids |b bcr-abl-directed antisense RNA and ribozymes |c Ralf Kronenwett, Rainer Haas, Georg Sczakiel |
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| 520 | |a Efficacy and sequence specificity are two major requirements in the use of antisense nucleic acids and ribozymes. For long-chain complementary RNA sequences (>30 nt), effects in living cells are correlated with the association rate of the complementary RNAin vitro, but not with the stability of the formed double strand. Thus, sequence selectivity of complementary RNA has to be defined as fastversusslow annealing with the appropriate target or non-target sequences, respectively. In this work, we performed a systematic kinetic analysis to evaluate the selectivity ofbcr-abl-directed antisense RNA and hammerhead ribozymes with a length of the complementary sequences of between 20 and 80 bases. By kineticin vitroselection, we identified oligomeric as well as long-chain complementary RNA that annealed at least tenfold faster with thebcr-ablsequence in comparison with either of the wild-type sequencesbcrorabl, respectively. In the presence of selected oligodeoxynucleotide sequences and RNase H, thebcr-abltranscript was specifically hydrolysed out of a mixture containingablandbcrsequences as well. Hammerhead ribozymes were designed such that binding with their target was facilitated eitherviahelix I or helix III-forming antisense arms but not both. Further, cleavage and binding occurred on opposite sides of thebcr-ablfusion point. Target selectivity was found for a ribozyme that annealed fastviaablsequences and cleaved within thebcrportion ofbcr-ablRNA. Kinetic probing and calculations of the local folding potential indicate that thebcr-ablfusion point sequences are not easily accessible for complementary nucleic acids. This study supports the need for more detailed structural investigations of thebcr-ablfusion sequence and forms a more rational basis for the therapeutic use of nucleic acid inhibitors of the aberrantbcr-ablgene expression in Philadelphia chromosome-positive cells. | ||
| 650 | 4 | |a antisense nucleic acids | |
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| 650 | 4 | |a RNA - RNA annealing | |
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