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An Alternative Culture Method to Maintain Genomic Hypomethylation of Mouse Embryonic Stem Cells Using MEK Inhibitor PD0325901 and Vitamin C
Published on: June 1, 2018
La vitamina C induce la desmetilación del ADN dependiente de Tet y un estado similar al blastocisto en las células ES
Kathryn Blaschke1, Kevin T Ebata, Mohammad M Karimi
1Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Department of Obstetrics and Gynecology and Center for Reproductive Sciences, University of California San Francisco, San Francisco, California 94143, USA.
Nature
|July 2, 2013
Resumen
La vitamina C mejora directamente la actividad de la enzima de translocación Ten eleven (Tet), aumentando los niveles de 5-hidroximetilcitocina (5hmC) y promoviendo la desmetilación del ADN en las células madre embrionarias de ratón, manteniendo así la fidelidad epigenética.
Área de la Ciencia:
- La epigenética es la epigenética.
- Biología Molecular Biología Molecular
- La genómica es la genómica.
Sus antecedentes:
- La metilación del ADN es una marca epigenética crucial que regula la expresión génica, la impresión y el silenciamiento del retrotransposón.
- Diez de once enzimas de translocación (Tet) median la desmetilación del ADN mediante la conversión de la 5-metilcitocina (5mC) en 5-hidroximetilcitocina (5hmC).
- Las células madre embrionarias de ratón (CEM) a menudo se cultivan sin vitamina C, un potencial cofactor para las enzimas Tet.
Objetivo del estudio:
- Investigar el papel de la vitamina C en la regulación de la actividad de la enzima Tet y la metilación del ADN en las ECS del ratón.
- Determinar si la vitamina C influye en la desmetilación global del ADN y la expresión génica en las ESC.
Principales métodos:
- Adición de vitamina C a las ECS de ratones de cultivo.
- Medición de los niveles de 5hmC y 5-metilcitosina (5mC).
- Análisis de la metilación del promotor genético y la expresión génica.
- Ensayos bioquímicos con el recombinante Tet1.1.
- Experimentos con el uso de Tet1 y Tet2 doble knockout ESCs.
Principales resultados:
- La vitamina C aumentó rápidamente y globalmente los niveles de 5hmC en los CES.
- Esto fue seguido por la desmetilación del ADN de los promotores de genes y la regulación al alza de los genes de la línea germinal.
- La vitamina C mejoró la actividad de Tet1 in vitro e in vivo, específicamente en regiones dependientes de Tet1/Tet2.
- El efecto de la vitamina C fue más pronunciado en las ganancias de metilación en los ESC cultivados en comparación con los blastocistos.
Conclusiones:
- La vitamina C actúa como un regulador directo de la actividad de la enzima Tet.
- La vitamina C es esencial para mantener la fidelidad de la metilación del ADN en las ESC.
- La vitamina C influye en la reprogramación epigenética durante el desarrollo temprano.
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