[Antisense oligonucleotides: tools of molecular genetics and therapeutic agents].
The binding of an oligodeoxynucleotide, so-called anti-sense, to the complementary sequence of a messenger RNA can prevent the synthesis of the encoded protein. This approach constitutes a very efficient and specific means to artificially regulate gene expression. Numerous chemical modifications have been introduced into synthetic oligos in order to provide them with properties that unmodified molecules do not display. For instance, oligos built up with methylphosphonate, phosphorothioate and alpha-anomer units lead to molecules that are resistant to DNases. Acridine-linked oligos exhibit an increased affinity for the target sequence due to the intercalation of the dye into the oligo/RNA duplex. Two different mechanisms account for translation inhibition by antisense oligos. Inhibition of the elongation step results only from the induced cleavage of the target RNA by RNase-H. In contrast, oligos targeted upstream of the AUG initiation codon can block the initiation step through an RNase-H independent mechanism. As a consequence, methylphosphonate- and alpha-oligos, which do not elicit RNase-H activity, targeted to the 5' region, are efficient antisense; but they are inactive if targeted to the coding sequence. Experiments performed with antisense oligos in cell-free extracts supported the notion that the mini-exon sequence, acquired by trans-splicing, was present on every message in trypanosomatids and on some of them in nematodes. Furthermore, an acridine-linked oligo complementary to the mini-exon sequence of Trypanosoma brucei induced a lethal effect on cultured procyclics. Therefore these compounds constitute promising tools in molecular genetics and could open new routes to rationally tailor therapeutic agents.
Toulmé
J J
JJ
Laboratoire de Biophysique, INSERM U201, CNRS UA481, Muséum National d'Histoire Naturelle, Paris, France.
Verspieren
P
P
Boiziau
C
C
Loreau
N
N
Cazenave
C
C
Thuong
N T
NT
D000818:Animals; D003851:Deoxyribonucleases / Q000378:metabolism; D005091:Exons; D015394:Molecular Structure; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000378:metabolism; D014346:Trypanosoma brucei brucei / Q000235:genetics*
D016376:Oligonucleotides, Antisense; D003851:Deoxyribonucleases
Les oligonucléotides antisens: outils de génétique moléculaire et agents thérapeutiques.