Lysine-specific Demethylase 1B (KDM1B) is a chromatin regulator which functions as a
histone eraser through the removal of the post-translational modifications mono and
dimethylation of histone 3 on lysine 4 (H3K4me1/2). This process is enhanced by the
formation of a complex with Nuclear Protein Glyoxylate Reductase (NPAC). NPAC resolves
the sequestration of the nucleosome histone tail to allow robust demethylation of
H3K4me2 by KDM1B, during transcriptional elongation by RNA polymerase 2 (RNAP II).
KDM1B is involved in many crucial processes during development. Its physiological
functions include the establishment of maternal genomic imprints, reset of the
epigenome during somatic cell reprogramming, and regulation of brown adipogenic
differentiation. In light of this, the role of KDM1B in human embryonic stem cells (hESCs)
is examined through CRISPR/Cas9-editing to further dissect its biological functions during
embryogenesis.
CRISPR-induced knockouts of KDM1B exhibited similar cell proliferation rate and
expression of OCT4 and NANOG pluripotency markers to wildtype cells. Furthermore,
KDM1B-/- clones were able to maintain their pluripotency potential by differentiating to
all germ layers in teratoma and embryoid body formation assays. In addition, RNA-seq of
KDM1B-/- clones showed enrichment of mesoderm lineage-related gene ontology (GO) terms in the downregulated differentially expressed genes. Thus, KDM1B is believed to
be dispensable during the pluripotent stage of the cell but proved fundamental during
later stages of development.
| Identifer | oai:union.ndltd.org:kaust.edu.sa/oai:repository.kaust.edu.sa:10754/662751 |
| Date | 04 1900 |
| Creators | Alfarhan, Dalal |
| Contributors | Adamo, Antonio, Biological and Environmental Science and Engineering (BESE) Division, Orlando, Valerio, Li, Mo |
| Source Sets | King Abdullah University of Science and Technology |
| Language | English |
| Detected Language | English |
| Type | Thesis |
| Rights | 2023-12-31, At the time of archiving, the student author of this thesis opted to temporarily restrict access to it. The full text of this thesis will become available to the public after the expiration of the embargo on 2023-12-31. |
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