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Series GSE125761 Query DataSets for GSE125761
Status Public on Jan 31, 2021
Title MYB is an essential regulator of primitive human hematopoiesis in human pluripotent stem cell differentiation cultures.
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Summary MYB is well recognized to be a key regulator of definitive hematopoiesis that plays an important role in the maintenance and multilineage differentiation of hematopoietic stem cells (HSCs). In the vertebrate developmental context, MYB is widely regarded dispensable for primitive hematopoiesis but critically required for the development of definitive hematopoiesis. To explore the role of MYB in human hematopoietic development we have inactivated the gene by bi-allelic TALEN-supported gene targeting in several lines of human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs), and subjected these cells to hematopoietic differentiation in well-defined cell culture conditions. Venus gene reporter was inserted into the knock-in allele to monitor MYB expression during the course of the hESC/iPSC differentiation. The gene reporter system showed that MYB is specifically expressed during hematopoietic commitment in the earliest primitive blood cells. Moreover, the level of MYB expression was highest at the commitment stage of differentiation and significantly decreased at the maturations stage. We found that MYB was not required for initial hematopoietic commitment of nascent mesoderm and emergence of primitive, yolk sac-type human hematopoietic progenitors. However, inactivation of MYB severely abrogated proliferation of the primitive erythroid and mixed erythroid-macrophage-megakaryocyte progenitors. In addition, MYB-negative hESC/iPSC lines demonstrated major defects in myeloid cell development and completely failed to generate mature granulocytes. Transposon-mediated rescue of MYB expression in MYB-null cells efficiently restored both the primitive hematopoietic progenitors and immature myeloid cells. Our data indicate that in contrast to its previously attributed exclusive role in definitive hematopoiesis, MYB is indispensable for primitive human hematopoiesis.
Overall design We inactivated the human MYB gene by biallelic integration of reporter donor cassette through TALENs-supported genetic engineering. The modified lines including MYB+/+, MYB+/- and MYB-/- were differentiated into hematopoietic cells in our new established serum-free, cytokine-free culture system with minimal BMP4 exposure for mesoderm induction. The cells were harvested at different time points for cell-sorting based on the expression of MYB-YFP and CD34 cell surface marker on day 6 and day 12. Subsequently the total mRNA was extracted using trizole reagent and subject to RNA-sequencing. In total 31 samples, 2 or 3 replicate samples per group being biological repeats, were analyzed.
Contributor(s) Shah Z, Samokhvalov IM
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Submission date Jan 28, 2019
Last update date Jan 31, 2021
Contact name ZAHIR SHAH
Phone 13229487422
Organization name GIBH,CAS, China
Department Stem Cells and Regenerative Medicines
Street address 190 Kaiyuan avenue, Haungpu district, Guangzhou, China
City Guangzhou
State/province Guangdong
ZIP/Postal code 510530
Country China
Platforms (2)
GPL16791 Illumina HiSeq 2500 (Homo sapiens)
GPL21290 Illumina HiSeq 3000 (Homo sapiens)
Samples (31)
GSM3581520 MYB-/-YFP-ve_day6rep1
GSM3581521 MYB-/-YFP-ve_day6rep2
GSM3581522 MYB-/-YFP+ve_day6rep1
BioProject PRJNA517462
SRA SRP182678

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Supplementary file Size Download File type/resource
GSE125761_RNAseqprocesseddata_description_v1.3.txt.gz 345 b (ftp)(http) TXT
GSE125761_genes_ntc_expression_cpmcounts_GEO.txt.gz 2.3 Mb (ftp)(http) TXT
GSE125761_genes_ntc_expression_rawcounts_GEO.txt.gz 1.8 Mb (ftp)(http) TXT
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