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Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids
Published on: September 21, 2017
El punto de unión y la secuencia de los conjugados peptídico-acridina inmovilizados controlan la afinidad por los
1Department of Chemistry, University of Utah, 315 South 1400 East, Salt Lake City, Utah 84112, USA.
Journal of the American Chemical Society
|July 18, 2002
Resumen
La inmovilización de los conjugados peptídico-acridina (PAC) para ensayos de unión de ácidos nucleicos requiere un apego estratégico. Tanto la secuencia conjugada como el punto de unión influyen en la afinidad por los ácidos nucleicos dúplex, lo que afecta a las estrategias de selección.
Área de la Ciencia:
- La bioquímica es la bioquímica.
- Biología Molecular Biología Molecular
- Biología Química Biología química.
Sus antecedentes:
- La inmovilización de fase sólida es crucial para la selección de bibliotecas combinatorias que utilizan arreglo espacial.
- La inmovilización de los intercaladores de ácido nucleico puede obstaculizar la unión si la funcionalización interfiere con el sitio de unión.
Objetivo del estudio:
- Desarrollar y evaluar un método para inmovilizar los conjugados peptídico-acridina (PAC) en soportes sólidos.
- Evaluar el impacto del sitio de inmovilización (terminal N o C) y la secuencia de aminoácidos en la afinidad de unión de ácidos nucleicos de los PAC.
Principales métodos:
- Los conjugados péptido-acridina (PAC) fueron sintetizados e inmovilizados a través del extremo C o N (posición 4 o 9 de la acridina).
- Se analizaron las propiedades de unión de ácido nucleico de los PAC inmovilizados.
- La afinidad para los ácidos nucleicos dúplex se determinó en función de la estrategia de inmovilización y la secuencia PAC.
Principales resultados:
- Tanto la secuencia de aminoácidos de la PAC como su punto de unión al soporte sólido afectan significativamente la afinidad de unión de los ácidos nucleicos.
- La inmovilización a través de diferentes terminales (4- frente a la 9-posición de la acridina) produce características de unión distintas.
- El estudio demuestra un método para analizar las propiedades de unión de los intercaladores inmovilizados.
Conclusiones:
- El diseño de los aglutinantes inmovilizados de ácido nucleico, en particular los intercaladores, debe tener en cuenta el punto de unión para evitar inhibir la unión.
- La inmovilización estratégica de los PAC es esencial para una selección efectiva de ligandos de ácido nucleico basada en microarrays y en perlas.
- Comprender la influencia del sitio de funcionalización es clave para desarrollar nuevas sondas y terapias moleculares basadas en intercaladores.
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