Development of a mouse embryonic stem cell model for investigating the functions of the linker histone H1-4.
Abed Alkarem Abu AlhaijaImtiaz Nisar LoneEsin Ozkuru SekerogluTugce BaturDimitar AngelovStefan DimitrovAli HamicheElif Nur Firat KaralarMuhammed Erdem ErcanTamer YagciHani AlotaibiMuhammed Kasim DirilPublished in: FEBS open bio (2024)
The linker histone H1 C-terminal domain (CTD) plays a pivotal role in chromatin condensation. De novo frameshift mutations within the CTD coding region of H1.4 have recently been reported to be associated with Rahman syndrome, a neurological disease that causes intellectual disability and overgrowth. To investigate the mechanisms and pathogenesis of Rahman syndrome, we developed a cellular model using murine embryonic stem cells (mESCs) and CRISPR/Cas9 genome engineering. Our engineered mES cells facilitate detailed investigations, such as H1-4 dynamics, immunoprecipitation, and nuclear localization; in addition, we tagged the mutant H1-4 with a photoactivatable GFP (PA-GFP) and an HA tag to facilitate pulldown assays. We anticipate that these engineered cells could also be used for the development of a mouse model to study the in vivo role of the H1-4 protein.
Keyphrases
- intellectual disability
- induced apoptosis
- crispr cas
- stem cells
- mouse model
- embryonic stem cells
- cell cycle arrest
- autism spectrum disorder
- genome wide
- genome editing
- gene expression
- case report
- endoplasmic reticulum stress
- oxidative stress
- signaling pathway
- high throughput
- dna methylation
- cell proliferation
- small molecule
- binding protein
- pi k akt
- brain injury
- wild type
- cerebral ischemia