Genome editing to model and reverse a prevalent mutation associated with myeloproliferative neoplasms
| Autor: | Ron Baik, Jacob E. Corn, Shaheen Kabir, Stacia K. Wyman |
|---|---|
| Přispěvatelé: | Bertolini, Francesco |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Cell signaling Heredity Cellular differentiation CD34 Artificial Gene Amplification and Extension Signal transduction Regenerative Medicine medicine.disease_cause Polymerase Chain Reaction Biochemistry Homozygosity Polycythemia vera 0302 clinical medicine Stem Cell Research - Nonembryonic - Human Animal Cells hemic and lymphatic diseases 2.1 Biological and endogenous factors Aetiology Post-Translational Modification Phosphorylation Cancer Erythroid Precursor Cells Gene Editing 0303 health sciences Mutation Heterozygosity Multidisciplinary Stem Cells Cell Differentiation Hematology 3. Good health STAT signaling Haematopoiesis 030220 oncology & carcinogenesis Medicine Stem Cell Research - Nonembryonic - Non-Human Cellular Types Stem cell Research Article General Science & Technology Science Biology Research and Analysis Methods Cell Line 03 medical and health sciences Rare Diseases Germline mutation Genetics medicine Humans Allele Progenitor cell Molecular Biology Techniques Molecular Biology 030304 developmental biology Myeloproliferative Disorders Wild type Biology and Life Sciences Proteins Cell Biology Janus Kinase 2 Stem Cell Research Hematopoietic Stem Cells medicine.disease Coculture Techniques 030104 developmental biology Cancer research CRISPR-Cas Systems Cloning Developmental Biology |
| Zdroj: | PLoS ONE, 16 (3) PLoS ONE PLoS ONE, Vol 16, Iss 3, p e0247858 (2021) PloS one, vol 16, iss 3 |
| ISSN: | 1932-6203 |
| Popis: | Myeloproliferative neoplasms (MPNs) cause the over-production of blood cells such as erythrocytes (polycythemia vera) or platelets (essential thrombocytosis). JAK2 V617F is the most prevalent somatic mutation in many MPNs, but previous modeling of this mutation in mice relied on transgenic overexpression and resulted in diverse phenotypes that were in some cases attributed to expression level. CRISPR-Cas9 engineering offers new possibilities to model and potentially cure genetically encoded disorders via precise modification of the endogenous locus in primary cells. Here we develop “scarless” Cas9-based reagents to create and reverse the JAK2 V617F mutation in an immortalized human erythroid progenitor cell line (HUDEP-2), CD34+ adult human hematopoietic stem and progenitor cells (HSPCs), and immunophenotypic long-term hematopoietic stem cells (LT-HSCs). We find no overt in vitro increase in proliferation associated with an endogenous JAK2 V617F allele, but co-culture with wild type cells unmasks a competitive growth advantage provided by the mutation. Acquisition of the V617F allele also promotes terminal differentiation of erythroid progenitors, even in the absence of hematopoietic cytokine signaling. Taken together, these data are consistent with the gradually progressive manifestation of MPNs and reveals that endogenously acquired JAK2 V617F mutations may yield more subtle phenotypes as compared to transgenic overexpression models. |
| Databáze: | OpenAIRE |
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