RNA sequencing comparing the effect of the introduction of CSF3R-d715 in control- or SCN- hematopoietic progenitor cells (HPCs)
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ABSTRACT: To address how CSF3R mutations affect hematopoiesis in a severe congenital neutropenia (SCN) background we used SCN patient, or control, derived induced pluripotent stem cells (iPSC). HPCs were generated from CRISPR-Cas9 genome edited CSF3R-d715 SCN or control-derived iPSCs with the STEMdiff™ Hematopoietic Kit (STEMCELL Technologies). Floating cells were harvested at Day 12 of the hematopoietic induction protocol. CD34+CD45+ HPCs were FACS sorted in TRIzol and RNA was isolated 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# 20015964), according to the low sample protocol and paired-end sequenced on a HiSeq 2500 or Novaseq 6000 (both Illumina).
INSTRUMENT(S): Illumina HiSeq 2500, Illumina NovaSeq 6000
ORGANISM(S): Homo sapiens
SUBMITTER: Patricia Olofsen
PROVIDER: E-MTAB-10288 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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