Los investigadores desarrollan una nueva herramienta computacional más precisa para la secuenciación de ARN de lectura larga

En su viaje del gen a la proteína, una molécula naciente de ARN puede cortarse y unirse, o empalmarse, de diferentes maneras antes de traducirse en una proteína. Este proceso, conocido como splicing alternativo, permite que un único gen codifique varias proteínas diferentes. El splicing alternativo se produce en muchos procesos biológicos, como cuando las células madre maduran y se convierten en células específicas de un tejido. Sin embargo, en el contexto de las enfermedades, el splicing alternativo puede estar desregulado. Por lo tanto, es importante examinar el transcriptoma -es decir, todas las moléculas de ARN que pueden derivarse de los genes- para comprender la causa fundamental de una enfermedad.

Sin embargo, históricamente ha sido difícil «leer» las moléculas de ARN en su totalidad porque suelen tener miles de bases. En su lugar, los investigadores han recurrido a la llamada secuenciación de ARN de lectura corta, que rompe las moléculas de ARN y las secuencia en trozos mucho más cortos -entre 200 y 600 bases, según la plataforma y el protocolo-. A continuación, se utilizan programas informáticos para reconstruir las secuencias completas de las moléculas de ARN.

Al encontrar áreas de coincidencias perfectas entre las lecturas de secuenciación de ARN largo y el ADN genómico, así como al tomar prestada información de todas las lecturas de secuenciación de ARN largo de un gen, la herramienta es capaz de identificar uniones de empalme e isoformas de ARN altamente fiables, incluidas aquellas que no se han documentado previamente en las bases de datos existentes.

Los investigadores evaluaron el rendimiento de ESPRESSO utilizando datos simulados y datos de muestras biológicas reales. Comprobaron que ESPRESSO funciona mejor que muchas de las herramientas disponibles en la actualidad, tanto a la hora de descubrir isoformas de ARN como de cuantificarlas. Los investigadores también generaron y analizaron más de mil millones de lecturas largas de secuenciación de ARN que cubrían 30 tipos de tejidos humanos y tres líneas celulares humanas, proporcionando un recurso útil para estudiar la variación del transcriptoma humano en la resolución de isoformas de ARN de longitud completa.

«ESPRESSO aborda un antiguo problema de secuenciación de ARN de larga lectura y podría abrir nuevas oportunidades de descubrimiento», afirmó el Dr. Xing. «Prevemos que ESPRESSO será una herramienta útil para que los investigadores exploren el repertorio de ARN de las células en diversos entornos biomédicos y clínicos».

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