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| Status |
Public on Mar 06, 2017 |
| Title |
Effect of BCL11B knockdown on transcriptome of human T-cell precursors |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
To investigate the role of BCL11B in the initial stages of human thymopoiesis, we performed loss of function (knockdown) studies in an in vitro human thymopoiesis model (cord blood CD34+ cells co-cultured on OP9DLL1 stromal cell line). Gene expression profiling by RNA-Seq demonstrated that BCL11B knockdown resulted in downregulation of T-lineage genes and upregulation of stem cell, myeloid and NK genes, indicating BCL11B is required for the establishment of a T-lineage commitment transcriptional program.
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| Overall design |
Cord blood CD34+ cells were transduced with a BCL11B knockdown or scrambled control shRNA lentivral vector containing a GFP reporter. GFP+ cells were isolated by fluorescence activation cell sorting (FACS) at 48 hours post transduction and cultured on OP9DLL1 stroma in the presence of FLT3 ligand (5 ng/ ml) and IL-7 (5 ng/ ml). On day 14 of co-culture, CD45+GFP+CD7+CD1a- and CD45+GFP+CD7+CD1a+ T-cell precursors were isolated from control and knockdown co-cultures by FACS and analyzed by RNA-Seq to determine the effect of BCL11B knockdown on the transcriptome of T-cell precursors. N=3 experiments, each done with a different pool of cord blood donors.
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| Contributor(s) |
Parekh C |
| Citation(s) |
28232744 |
| |
| Submission date |
Jan 21, 2017 |
| Last update date |
May 15, 2019 |
| Contact name |
chintan parekh |
| E-mail(s) |
[email protected]
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| Organization name |
Children's Hospital Los Angeles
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| Department |
Pediatrics
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| Street address |
4650 sunset blvd, mail stop 54
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| City |
Los Angeles |
| State/province |
California |
| ZIP/Postal code |
90027 |
| Country |
USA |
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| Platforms (1) |
| GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
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| Samples (12)
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| This SubSeries is part of SuperSeries: |
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| Relations |
| BioProject |
PRJNA362787 |
| SRA |
SRP097469 |
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