Seleção e caracterização de aptâmeros de RNA ligantes a regiao 5’-UTR do genoma do virus da dengue
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Data
2014-01-21
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Universidade Federal de Goiás
Resumo
The increasing number of notifications of dengue infections is becoming a very
important concern for global healthcare programs. Combinatorial technologies
aiming the selection of specific short conformational nucleic acid ligands against
viral targets, also called aptamers, can be achieved by large-scale selections
using the genomic SELEX technology. Our hypothesis is that aptamers can be
directly selected against dengue RNA conformational structures that present
functional elements in the 5'-UTR sequence, which form RNA-RNA and protein-
RNA interactions, and play significant roles in the infection process. Our aim
was to select and characterize aptamers that bind to the 5’-UTR using the
matrix-free SELEX method. Products from the eighth selection cycle were
isolated, cloned and sequenced, and 14 ligands were chosen for in
silico characterization. Aptamers were grouped into three families according to
their sequence homology, and conserved ribonucleotides generated specific
linear motifs. Sequences motifs were detected in random nucleotides regions
ranging from 31 to 40 nt, which showed higher affinity to DENV1 and 3 virus.
The novel molecules and processes described in this study open new insights
for dengue research and applications, and the selected aptamers can be used
either as diagnostic or therapeutic tools. In silico analyses revealed that
aptamer binding to its RNA target may lead to alterations of viral RNA
secondary structures, and is probably leading to the loss of its original
conformational and preventing its replication and/or the transcription
process. The analyses also demonstrated that aptamers presented a broad
hybridization spectrum to DENV1 and 3 even in the presence of mutations in
different subtypes, which suggest its possible use in the other two serotypes,
DENV2 and 4. This is the first description of aptamers against the RNA
structure of Dengue Virus with important implications in the disease control.
Descrição
Palavras-chave
Dengue, SELEX, Aptâmeros, Ribonucleotídeo, Aptamers, Ribonucleotide
Citação
CNOSSEN, Elismar de Jesus Nunes. Seleção e caracterização de aptâmeros de RNA ligantes a regiao 5’-UTR do genoma do virus da dengue. 2014. 71 f. Dissertação (Mestrado em Química) - Universidade Federal de Goiás, Catalão, 2014.