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Cruciform Formable Sequences within Pou5f1 Enhancer Are Indispensable for Mouse ES Cell Integrity
Yu Yamamoto1,2, Osamu Miura2, Takashi Ohyama1,2
1Department of Biology, Faculty of Education and Integrated Arts and Sciences, Waseda University, Shinjuku-ku, Tokyo 162-8480, Japan.
International Journal of Molecular Sciences
|April 3, 2021
Summary
Cruciform-forming inverted repeats (CFIRs) in the mouse Pou5f1 enhancer regulate pluripotency genes. Altering CFIRs impacts Pou5f1, Sox2, Nanog, Klf4, and Esrrb expression, suggesting a role in maintaining embryonic stem cell integrity.
Area of Science:
- Molecular Biology
- Genetics
- Epigenetics
Background:
- DNA adopts various structures beyond the canonical B-form, including cruciform structures formed by inverted repeat (IR) sequences.
- Cruciform-forming IRs (CFIRs) are found in transcriptional regulatory regions, but their functional significance in eukaryotic transcription remains largely unknown.
Purpose of the Study:
- To investigate the role of CFIRs within the mouse Pou5f1 enhancer in transcriptional regulation.
- To elucidate the in vivo function of a cluster of CFIRs in regulating pluripotency genes and non-coding RNAs.
Main Methods:
- Identification of a CFIR cluster within the mouse Pou5f1 enhancer.
- Genome editing in mouse embryonic stem (ES) cells to convert CFIRs to mirror repeat sequences.
- Quantitative analysis of transcript levels for Pou5f1, Sox2, Nanog, Klf4, Esrrb, and associated non-coding RNAs.
Main Results:
- Alteration of CFIRs in mouse ES cells led to decreased Pou5f1 transcript levels.
- Genome editing resulted in elevated expression of Sox2, Nanog, Klf4, and Esrrb transcripts.
- Transcription of non-coding RNAs within the enhancer was upregulated in genome-edited cells, correlating with the expression changes of the pluripotency genes.
Conclusions:
- The CFIRs within the Pou5f1 enhancer play an active role in the transcriptional regulation of key pluripotency genes.
- These CFIRs, potentially through associated non-coding RNAs, are crucial for maintaining the integrity and function of mouse embryonic stem cells.
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