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Atlas de la localización del ARN subcelular revelado por APEX-Seq

Furqan M Fazal¹, Shuo Han², Kevin R Parker¹

¹Center for Personal Dynamics Regulomes, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Dermatology, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.

|June 25, 2019

Resumen

Desarrollamos APEX-seq, un nuevo método de secuenciación de ARN que mapea el transcriptoma espacial con resolución nanométrica. Esta técnica revela patrones de localización de ARN dentro de las células, incluida la organización nuclear y el transporte de ARNm mitocondrial.

Área de la Ciencia:

Biología molecular
La genómica
Biología celular

Sus antecedentes:

La comprensión de la localización del ARN es crucial para la regulación de la expresión génica.
Los métodos existentes carecen de la resolución para mapear la distribución de transcripciones dentro de los compartimentos subcelulares.

Objetivo del estudio:

Para introducir APEX-seq, un método de secuenciación de ARN dependiente de la proximidad.
Para crear un mapa espacial de alta resolución del transcriptoma humano.
Para investigar los patrones de localización de ARN en distintas localizaciones subcelulares.

Principales métodos:

Utilizó la enzima peroxidasa APEX2 para el etiquetado de proximidad directa del ARN.
Aplicamos APEX-seq en nueve ubicaciones subcelulares distintas.
Datos de transcriptomas espaciales generados con resolución nanométrica.

Principales resultados:

Reveló extensos patrones de localización para diversas clases de ARN y isoformas de transcripción.
Descubrió una organización radial del transcriptoma nuclear.
Se identificaron distintas vías de localización del ARN mensajero a las mitocondrias.

Conclusiones:

APEX-seq proporciona una poderosa herramienta para mapear el transcriptoma espacial.
El método permite investigaciones exhaustivas de la localización del ARN en varios sistemas biológicos.
Los hallazgos ofrecen información sobre la exportación nuclear y la regulación de la expresión génica mitocondrial.

Palabras clave:

Los LAD OXPHOS también Las UTR cicloheximida Los motivos Nocodazol y sus derivados Los retrotransposones Transcriptómica espacial y sus aplicaciones Traducción

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