The Pu.1 target gene Zbtb11 regulates neutrophil development through its integrase-like HHCC zinc finger

From a forward genetic screen in zebrafish, we identified the transcription factor, ZBTB11, as critical for basal and emergency granulopoiesis and showed that ZBTB11 sits in a pathway directly downstream of master myeloid regulators including PU.1, GFI1 and CEBPa. To better understand target genes regulated by Zbtb11, RNAseq profiling was performed in neutrophils from WT and Zbtb11 mutant zebrafish. At 48 hours post fertilisation, single-cell suspensions were prepared from either pooled WT embryos or from pooled phenotype-sorted zbtb11 mutant embryos. Neutrophils were FACS sorted on the basis of bright fluorescence for the transgenic neutrophil markers mpx:EGFP or lyz:dsRed into RNALater. N=3 biologically independent pools of neutrophils were obtained from WT and from zbtb11 mutant embryos and used for preparation of RNA and sequencing libraries.