Locus-Specific Isolation of the Nanog Chromatin Identifies Regulators Relevant to Pluripotency of Mouse Embryonic Stem Cells and Reprogramming of Somatic Cells
Abstract
:1. Introduction
2. Results
2.1. CAPTURE of the Nanog Promoter
2.2. Isolation of Proteins Assembled on the Endogenous Nanog Chromatin
2.3. Effects of CAPTURE-Identified Proteins on mESC Pluripotency
2.4. Roles for the Identified Proteins in mESC Differentiation
2.5. Roles for the Identified Proteins in Somatic Cell Reprogramming
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Plasmid Construction
4.3. Production and Infection of Viral Vectors
4.4. CAPTURE of the Nanog Promoter
4.4.1. Isolation of Chromatin Containing the Nanog Promoter
4.4.2. CAPTURE-ChIP-qPCR
4.4.3. CAPTURE-Proteomics
4.4.4. Gene Ontology Analysis
4.5. Genome Editing
4.6. Cell Sorting and Image Analysis
4.7. Gene Expression Analysis
4.8. Somatic Cell Reprogramming
4.9. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Burramsetty, A.K.; Nishimura, K.; Kishimoto, T.; Hamzah, M.; Kuno, A.; Fukuda, A.; Hisatake, K. Locus-Specific Isolation of the Nanog Chromatin Identifies Regulators Relevant to Pluripotency of Mouse Embryonic Stem Cells and Reprogramming of Somatic Cells. Int. J. Mol. Sci. 2022, 23, 15242. https://doi.org/10.3390/ijms232315242
Burramsetty AK, Nishimura K, Kishimoto T, Hamzah M, Kuno A, Fukuda A, Hisatake K. Locus-Specific Isolation of the Nanog Chromatin Identifies Regulators Relevant to Pluripotency of Mouse Embryonic Stem Cells and Reprogramming of Somatic Cells. International Journal of Molecular Sciences. 2022; 23(23):15242. https://doi.org/10.3390/ijms232315242
Chicago/Turabian StyleBurramsetty, Arun Kumar, Ken Nishimura, Takumi Kishimoto, Muhammad Hamzah, Akihiro Kuno, Aya Fukuda, and Koji Hisatake. 2022. "Locus-Specific Isolation of the Nanog Chromatin Identifies Regulators Relevant to Pluripotency of Mouse Embryonic Stem Cells and Reprogramming of Somatic Cells" International Journal of Molecular Sciences 23, no. 23: 15242. https://doi.org/10.3390/ijms232315242