RNA sequencing comparing the effect of the introduction of RUNX1-D171N in Csf3r-d715 mice bone marrow cells

ABSTRACT: To address how Csf3r and RUNX1 mutations in combination with CSF3 administration affect hematopoiesis in vitro, we performed hematopoietic expansion cultures in LODISH stem cell expansion medium. Lineage-negative Csfr-d715 BM cell cells were retrovirally transduced with the patient specific RUNX1-D171N (RHD) mutant or an empty vector control and subsequently treated with puromycin to select for the transduced cells. Lineage- c-Kit+ (LK) cells were FACS sorted in TRIzol 2, 5 and 9 days after puromycin selection and subsequently used for RNA isolation according to the manufacturer’s protocol. SMARTer Ultra Low Input RNA kit for sequencing (Clontech, v4 Cat# 634891) was used to generate cDNA. Sequencing libraries were generated using TruSeq Nano DNA Sample Preparation kits (Illumina, Cat# 200

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Mus musculus

SUBMITTER: Patricia Olofsen 

PROVIDER: E-MTAB-9373 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress