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Impaired stem cell differentiation and somatic cell reprogramming in DIDO3 mutants with altered RNA processing and increased R-loop levels

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2021

Cell Death Dis. 2021 Jun 21;12(7):637. doi: 10.1038/s41419-021-03906-2.

Impaired stem cell differentiation and somatic cell reprogramming in DIDO3 mutants with altered RNA processing and increased R-loop levels

Agnes Fütterer, Amaia Talavera-Gutiérrez, Tirso Pons, Jesús de Celis, Julio Gutiérrez, Verónica Domínguez Plaza, Carlos Martínez-A

Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB-CSIC), 28049, Madrid, Spain. Transgenesis Unit, CNB & Centro de Biología Molecular Severo Ochoa, CSIC-Universidad Autónoma de Madrid, 28049, Madrid, Spain. Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB-CSIC), 28049, Madrid, Spain.

Service type: Knockout mice

Abstract

Embryonic stem cell (ESC) differentiation and somatic cell reprogramming are biological processes governed by antagonistic expression or repression of a largely common set of genes. Accurate regulation of gene expression is thus essential for both processes, and alterations in RNA processing are predicted to negatively affect both. We show that truncation of the DIDO gene alters RNA splicing and transcription termination in ESC and mouse embryo fibroblasts (MEF), which affects genes involved in both differentiation and reprogramming. We combined transcriptomic, protein interaction, and cellular studies to identify the underlying molecular mechanism. We found that DIDO3 interacts with the helicase DHX9, which is involved in R-loop processing and transcription termination, and that DIDO3-exon16 deletion increases nuclear R-loop content and causes DNA replication stress. Overall, these defects result in failure of ESC to differentiate and of MEF to be reprogrammed. MEF immortalization restored impaired reprogramming capacity. We conclude that DIDO3 has essential functions in ESC differentiation and somatic cell reprogramming by supporting accurate RNA metabolism, with its exon16-encoded domain playing the main role.

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