Relative contribution of photo-addition, helper oligonucleotide and RNase H to the antisense effect of psoralen-oligonucleotide conjugates, on in vitro translation of Leishmania mRNAs
Identifieur interne : 001E13 ( Istex/Curation ); précédent : 001E12; suivant : 001E14Relative contribution of photo-addition, helper oligonucleotide and RNase H to the antisense effect of psoralen-oligonucleotide conjugates, on in vitro translation of Leishmania mRNAs
Auteurs : Emanuelle Pascolo [France] ; Denis Hudrisier [France] ; Brian Sproat [Allemagne] ; Nguyen T. Thuong [France] ; Jean-Jacques Toulme [France]Source :
- BBA - Gene Structure and Expression [ 0167-4781 ] ; 1994.
English descriptors
- Teeft :
- Acad, Acta, Amazonensis, Antisense, Antisense effect, Antisense oligonucleotide, Antisense oligonucleotides, Base pairs, Biochimica, Biophysica, Biophysica acta, Denaturing polyacrylamide, Duplex, Electrophoretic analysis, Gene expression, Hairpin, Hairpin structure, Helper oligonucleotide, Leishmania, Leishmania amazonensis, Leishmania mrnas, Mrna, Natl, Nucleic, Nucleic acids, Oligomer, Oligomers, Oligonucleotide, Oligonucleotides, Pascolo, Polyacrylamide, Previous study, Proc, Protein synthesis, Psoralen, Psoralen derivatives, Rabbit reticulocyte lysate, Reticulocyte, Rnase, Simultaneous addition, Thuong, Toulm, Toulm6, Wheat germ.
Abstract
Abstract: We investigated the properties of two antisense oligonucleotides, 11αPso and 14TMP, 11 and 14 nucleotides long, respectively, and conjugated to psoralen derivatives. These oligonucleotides were complementary to the mini-exon sequence of Leishmania amazonensis. Upon ultraviolet (UV) irradiation these oligomers were selectively cross-linked to DNA or RNA target sequences, either 14 or 35 nucleotides long. The yield of photo-addition was much lower on the longer targets than on the shorter ones, due to the presence of a hairpin structure. The co-addition of a helper oligonucleotide, whose binding site, on the 35-mer, was adjacent to that of the psoralen-derivatized antisense oligomer, improved the cross-linking efficiency. We then determined the effect of 14TMP on in vitro translation of Leishmania mRNA in cell-free extracts. Non-irradiated antisense oligonucleotide/mRNA complexes reduced the protein synthesis in wheat germ extract but not in rabbit reticulocyte lysate. Conversely, UV irradiation induced a 14TMP-dependent reduction of translation in reticulocyte lysate whereas the inhibition was not improved in the wheat germ extract. These results are discussed with respect to the involvement of RNase-H in the oligonucleotide-mediated effect on protein synthesis.
Url:
DOI: 10.1016/0167-4781(94)90251-8
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<term>Acta</term>
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<term>Antisense</term>
<term>Antisense effect</term>
<term>Antisense oligonucleotide</term>
<term>Antisense oligonucleotides</term>
<term>Base pairs</term>
<term>Biochimica</term>
<term>Biophysica</term>
<term>Biophysica acta</term>
<term>Denaturing polyacrylamide</term>
<term>Duplex</term>
<term>Electrophoretic analysis</term>
<term>Gene expression</term>
<term>Hairpin</term>
<term>Hairpin structure</term>
<term>Helper oligonucleotide</term>
<term>Leishmania</term>
<term>Leishmania amazonensis</term>
<term>Leishmania mrnas</term>
<term>Mrna</term>
<term>Natl</term>
<term>Nucleic</term>
<term>Nucleic acids</term>
<term>Oligomer</term>
<term>Oligomers</term>
<term>Oligonucleotide</term>
<term>Oligonucleotides</term>
<term>Pascolo</term>
<term>Polyacrylamide</term>
<term>Previous study</term>
<term>Proc</term>
<term>Protein synthesis</term>
<term>Psoralen</term>
<term>Psoralen derivatives</term>
<term>Rabbit reticulocyte lysate</term>
<term>Reticulocyte</term>
<term>Rnase</term>
<term>Simultaneous addition</term>
<term>Thuong</term>
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<front><div type="abstract" xml:lang="en">Abstract: We investigated the properties of two antisense oligonucleotides, 11αPso and 14TMP, 11 and 14 nucleotides long, respectively, and conjugated to psoralen derivatives. These oligonucleotides were complementary to the mini-exon sequence of Leishmania amazonensis. Upon ultraviolet (UV) irradiation these oligomers were selectively cross-linked to DNA or RNA target sequences, either 14 or 35 nucleotides long. The yield of photo-addition was much lower on the longer targets than on the shorter ones, due to the presence of a hairpin structure. The co-addition of a helper oligonucleotide, whose binding site, on the 35-mer, was adjacent to that of the psoralen-derivatized antisense oligomer, improved the cross-linking efficiency. We then determined the effect of 14TMP on in vitro translation of Leishmania mRNA in cell-free extracts. Non-irradiated antisense oligonucleotide/mRNA complexes reduced the protein synthesis in wheat germ extract but not in rabbit reticulocyte lysate. Conversely, UV irradiation induced a 14TMP-dependent reduction of translation in reticulocyte lysate whereas the inhibition was not improved in the wheat germ extract. These results are discussed with respect to the involvement of RNase-H in the oligonucleotide-mediated effect on protein synthesis.</div>
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