Project description:We FACS purified GFP+ cells 2 days post-transduction of LSK (Lin-Negative c-Kit+ Sca1+) cells with miR-99 KD and Scr lentiviral vectors and performed RNA-sequencing; this allowed us to identify potential miR-99 target genes for inclusion in the shRNA library
Project description:We FACS-purified GFP+ LSK cells from mice transplanted with stably engrafted miR-99 KD and Scr HSCs and performed RNA-sequencing, demonstrating miR-99 KD results in significant depletion of hematopoietic stem cell gene expression signature and induces a differentiated progenitor gene expression signature.
Project description:We FACS purified L-GMP cells (GFP+ tdTom+ c-Kit+ Sca-1+ CD16/32+ CD34+) from mice transplanted with post-transformed Scr or miR-99 KD MLL-AF9 bone marrow and performed RNA-sequencing, demonstrating miR-99 KD results in induction differentiation and enrichment for a normal GMP gene expression signature
Project description:Long-term hematopoietic stem cells (LT-HSC) were sorted from human cord blood and cultured overnight before transduction with lentiviral vectors (empty E CTRL or MYC). Six days after transduction, BFP+ transduced cells were sorted for RNA extraction and sequencing
Project description:RNAseq analysis of USP7 shRNA KD in T-ALL cell lines. USP7 shRNAs lentivirus was generated by co-transfecting 293T cells with shRNA vectors (OriGene or Sigma). T-ALL cells were transduced with USP7 shRNA lentivirus and sorted for GFP positive cells five days after transduction or selected by puromycin.
Project description:Long-term hematopoietic stem cells (LT-HSC) were sorted from human cord blood and cultured overnight before transduction with lentiviral vectors overexpressing an shRNA against Renilla (shCTRL) or TFEB (shTFEB). Six days later, mCherry+ transduced cells were sorted for RNA extraction and sequencing.
Project description:We performed CUT&RUN assay with control Ig , anti-Flag and anti-ETO antibodies in Flag-tag AML1-ETO Kasumi-1 cells to determine binding occupancy of AML1-ETO following miR-130a KD. Kasumi-1 cells were transduced in 3 independent replicates for control and miR-130a KD samples with lentiviruses containing GFP reporter gene. GFP+ cells were sorted 3 days post-transduction and the cells were used in the CUT&RUN assay. Three antibodies were used for each biological sample.
Project description:Mouse LT-HSC were sorted and cultured in mScf, mTpo, mFlt3L, hIGFBP2 and Angptl5 for 2 days. These expression values were related to insertions of gamma-retroviral, lentiviral or alpharetroviral vectors carrying GFP which were retrieved after serial murine BM transplantation. The relation between gene expression in the cells responsible for long-term hematopoiesis and location of vector integration was investigated. Biological duplicate measurements of murine sorted and cultured LT-HSC (LSK, CD34-, CD135-)
Project description:In this experiment, we want to assess the effect of a lentiviral miR-10a and miR-335 overexpression on the transcriptome of murine LSK (Lin-,Sca-1+,c-Kit+) cells. Primary LSK cells were transduced with lentiviral miRNA overexpression constructs (control: GFP overexpression) and sorted for transduced cells (GFP+) after five days of in vitro culture (Flt-3, TPO, IL-3, SCF containing media).
Project description:To investigate the molecular and biological pathways altered by S1PR3OE in human hematopoietic stem cells (HSC), we performed RNA-sequencing (RNA-seq) of LT- and ST-HSC 3 days after transduction with control or S1PR3 overexpression (OE) lentiviral vectors
