Selection and characterization of active hammerhead ribozymes targeted against cyclin E and E2F1 full-length mRNA.
Antisense Nucleic Acid Drug Dev
; 11(5): 271-87, 2001 Oct.
Article
en En
| MEDLINE
| ID: mdl-11763345
Proliferation of vascular smooth muscle cells is generally accepted as a key event in the development of restenosis following percutaneous transluminal angioplasty. To prevent human restenosis, we have designed a molecular strategy based on hammerhead ribozymes targeted against the mRNA of cyclin E and E2F1, two proteins relevant in cell cycle progression whose regulation is interconnected by a positive feedback loop. Following the identification of accessible ribozyme target sites by RNase H mapping, several hammerhead ribozymes were generated that cleave with comparable efficiency two different splice forms of cyclin E mRNA and the full-length and a truncated form of E2F1 RNA, respectively. The most active ribozymes were tested in vitro under single-turnover conditions yielding k(react)/K(m) ratios between 36 and 73 x 10(4) M(-1) min(-1), which places them in the top range ribozymes targeted against long and structured substrates. In addition, we show that the most active ribozyme selected in vitro reduces specifically and significantly (p < 0.0028) proliferation of cultured human vascular smooth muscle cells (VSMC).
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Factores de Transcripción
/
ARN Mensajero
/
ARN Catalítico
/
Proteínas de Ciclo Celular
/
Ciclina E
/
Proteínas de Unión al ADN
Tipo de estudio:
Prognostic_studies
Límite:
Humans
Idioma:
En
Revista:
Antisense Nucleic Acid Drug Dev
Asunto de la revista:
BIOLOGIA MOLECULAR
/
BIOQUIMICA
/
FARMACOLOGIA
Año:
2001
Tipo del documento:
Article
País de afiliación:
Alemania
Pais de publicación:
Estados Unidos