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1.
Nucleic Acids Res ; 47(14): 7580-7591, 2019 08 22.
Artículo en Inglés | MEDLINE | ID: mdl-31147722

RESUMEN

Alternative cleavage and polyadenylation (APA) can occur at more than half of all human genes, greatly enhancing the cellular repertoire of mRNA isoforms. As these isoforms can have altered stability, localisation and coding potential, deregulation of APA can disrupt gene expression and this has been linked to many diseases including cancer progression. How APA generates cancer-specific isoform profiles and what their physiological consequences are, however, is largely unclear. Here we use a subcellular fractionation approach to determine the nuclear and cytoplasmic APA profiles of successive stages of colon cancer using a cell line-based model. Using this approach, we show that during cancer progression specific APA profiles are established. We identify that overexpression of hnRNPC has a critical role in the establishment of APA profiles characteristic for metastatic colon cancer cells, by regulating poly(A) site selection in a subset of genes that have been implicated in cancer progression including MTHFD1L.


Asunto(s)
Empalme Alternativo , Ribonucleoproteína Heterogénea-Nuclear Grupo C/genética , Neoplasias/genética , Poliadenilación , Isoformas de ARN/genética , Aminohidrolasas/genética , Aminohidrolasas/metabolismo , Línea Celular Transformada , Línea Celular Tumoral , Neoplasias del Colon/genética , Neoplasias del Colon/metabolismo , Neoplasias del Colon/patología , Progresión de la Enfermedad , Formiato-Tetrahidrofolato Ligasa/genética , Formiato-Tetrahidrofolato Ligasa/metabolismo , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Ribonucleoproteína Heterogénea-Nuclear Grupo C/metabolismo , Humanos , Metilenotetrahidrofolato Deshidrogenasa (NADP)/genética , Metilenotetrahidrofolato Deshidrogenasa (NADP)/metabolismo , Complejos Multienzimáticos/genética , Complejos Multienzimáticos/metabolismo , Neoplasias/metabolismo , Neoplasias/patología , Poli A/genética , Poli A/metabolismo , Interferencia de ARN
2.
Crit Rev Biotechnol ; 40(2): 265-281, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-31928250

RESUMEN

As a host for therapeutic protein expression, Chinese hamster ovary (CHO) cells are widely utilized in the mainstream biopharmaceutical industry. Cell culture process development plays an important role in transitioning laboratory research to manufacturing. Among different mathematic tools, kinetic modeling is commonly achieved through analyzing cell culture data to design process parameters, optimize media, and scale up bioreactors. In this review, we examine key factors for upstream process development, and summarize currently used kinetic modeling strategies. In addition, two original examples of kinetic modeling application optimizing cell culture performance are presented. A comprehensive understanding is provided for the kinetic modeling and its applications in cell culture process development.


Asunto(s)
Reactores Biológicos , Proteómica , Animales , Células CHO , Técnicas de Cultivo de Célula , Cricetinae , Cricetulus , Cinética
3.
RNA Biol ; 14(7): 865-890, 2017 07 03.
Artículo en Inglés | MEDLINE | ID: mdl-28453393

RESUMEN

Cleavage and polyadenylation (pA) is a fundamental step that is required for the maturation of primary protein encoding transcripts into functional mRNAs that can be exported from the nucleus and translated in the cytoplasm. 3'end processing is dependent on the assembly of a multiprotein processing complex on the pA signals that reside in the pre-mRNAs. Most eukaryotic genes have multiple pA signals, resulting in alternative cleavage and polyadenylation (APA), a widespread phenomenon that is important to establish cell state and cell type specific transcriptomes. Here, we review how pA sites are recognized and comprehensively summarize how APA is regulated and creates mRNA isoform profiles that are characteristic for cell types, tissues, cellular states and disease.


Asunto(s)
Poliadenilación/genética , ARN Mensajero/metabolismo , Animales , Humanos , Modelos Biológicos , Caperuzas de ARN/metabolismo , División del ARN/genética , Empalme del ARN/genética , ARN Mensajero/genética
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