Programming a highly structured ribozyme into complex allostery using RNA oligonucleotides.
ACS Chem Biol
; 7(11): 1802-6, 2012 Nov 16.
Article
em En
| MEDLINE
| ID: mdl-22950875
ABSTRACT
RNA possesses great potential for expanding the toolbox currently available to synthetic biologists. Here, the modulation of the Hepatitis Delta Virus ribozyme's activity with a series of rationally designed aptamers and effector RNA oligonucleotides is described. The ribozyme was initially fused with an 18-nucleotide hairpin structure that abolished its self-cleaving activity. The binding of a 14-mer oligonucleotide to the hairpin rescued the self-cleavage in a concentration-dependent manner. This modified ribozyme was inserted into the 5' UTR of a reporter gene, and the resulting construct was used to demonstrate that it is possible to modulate the ribozyme activity in cellulo with the oligonucleotide. Subsequently, ribozymes possessing specialized aptamers respecting other logic gates were also successfully designed and found to be functional in vitro. To our knowledge, this is the first example of HDV ribozyme regulation by oligonucleotides, as well as the first allosteric regulation of HDV ribozyme in mammalian cells.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Vírus Delta da Hepatite
/
RNA Catalítico
/
Aptâmeros de Nucleotídeos
Limite:
Humans
Idioma:
En
Revista:
ACS Chem Biol
Ano de publicação:
2012
Tipo de documento:
Article