A Genome-wide CRISPR screen to identify novel pathways involved in reprogramming and x-chromosome reactivation

During reprogramming of somatic cells into induced pluripotent stem cells (iPSCs), the epigenome needs to be reset. A prime example of this is the reversal of the silent state of the inactive X chromosome in female cells, which is achieved in a process called X-chromosome reactivation (XCR). Few pla...

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Detalles Bibliográficos
Autor: Barrero Asencio, Mercedes
Tipo de recurso: tesis doctoral
Estado:Versión publicada
Fecha de publicación:2023
País:España
Institución:CBUC, CESCA
Repositorio:TDR. Tesis Doctorales en Red
OAI Identifier:oai:www.tdx.cat:10803/687840
Acceso en línea:http://hdl.handle.net/10803/687840
Access Level:acceso abierto
Palabra clave:X chromosome
Reprogramming
Pluripotency
CRISPR screen
Interferon γ
Cromosoma X
Reprogramación
Pluripotencia
Cribado de CRISPR
Interferón γ
577
Descripción
Sumario:During reprogramming of somatic cells into induced pluripotent stem cells (iPSCs), the epigenome needs to be reset. A prime example of this is the reversal of the silent state of the inactive X chromosome in female cells, which is achieved in a process called X-chromosome reactivation (XCR). Few players have been described to be involved in XCR so far, and a comprehensive understanding of the regulatory networks has been lacking. Therefore, in this doctoral thesis I aim to shed light onto the mechanism of XCR during somatic cell reprogramming into iPSCs, by performing a genome-wide CRISPR-screen during this process. Using this approach, I identified a number of previously known and unknown pathways which are involved in reprogramming and/or XCR. Of these, I focused on the interferon γ (IFNγ) pathway, the activation of which during the early phase of reprogramming accelerated pluripotency acquisition and XCR. In this study, I sought to uncover the mechanism by which IFNγ enhances XCR, and found a dependency on TET-enzyme activity and DNA demethylation. These findings will contribute to the mechanistic understanding of the process of XCR and could have a potential impact on improving iPSC generation.