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Detection of Alternative Splicing During Epithelial-Mesenchymal Transition
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Developmental Xist induction is mediated by enhanced splicing.

Cheryl Stork1, Zhelin Li2, Lin Lin3

  • 1Graduate Program in Cell, Molecular and Developmental Biology, University of California, Riverside, Riverside, CA 92521, USA.

Nucleic Acids Research
|November 30, 2018
PubMed
Summary
This summary is machine-generated.

X-chromosome inactivation relies on Xist RNA splicing. Inefficient splicing in embryonic stem cells prevents Xist activity, acting as a crucial fail-safe mechanism during development.

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Area of Science:

  • Epigenetics and Gene Regulation
  • RNA Biology
  • Mammalian Development

Background:

  • X-inactive-specific transcript (Xist) is a long noncoding RNA (lncRNA) critical for X chromosome inactivation in female mammals.
  • Xist RNA mediates the silencing of one X chromosome, a process essential for dosage compensation.
  • The regulation of Xist RNA maturation, particularly splicing, is key to its function.

Purpose of the Study:

  • To investigate the role of Xist RNA splicing in the regulation of X chromosome inactivation.
  • To determine how Xist splicing efficiency changes during embryonic stem cell differentiation.
  • To explore the impact of splicing factors, like polypyrimidine tract binding protein 1 (Ptbp1), on Xist RNA maturation.

Main Methods:

  • Analysis of Xist RNA splicing status in naive and differentiated embryonic stem cells (ESCs).
  • Investigation of Xist splicing in genetically modified cells, including those with mutated Ptbp1.
  • Comparative analysis of Xist splicing efficiency between different Xist alleles (e.g., Xist129 and XistCAS) in hybrid ESCs.
  • Single-cell analysis to link Xist splicing choice to X chromosome inactivation.

Main Results:

  • Xist RNA is predominantly unspliced in naive ES cells and becomes efficiently spliced upon differentiation.
  • Defects in Xist splicing, caused by mutations in Ptbp1, impair the upregulation of mature Xist RNA.
  • Allelic differences in Xist splicing efficiency (Xist129 vs. XistCAS) correlate with preferential X chromosome inactivation.
  • Single-cell data reveal that Xist splicing choice is associated with the inactive X chromosome.

Conclusions:

  • Post-transcriptional control of Xist RNA splicing is a critical regulatory step for Xist induction and function.
  • Inefficient Xist splicing in ES cells serves as a fail-safe mechanism to prevent premature X chromosome inactivation.
  • Splicing plays a significant role in the developmental regulation of nuclear-retained lncRNAs like Xist.