Effect of MKRN3 deletion on gene expression in hypothalamic neurons derived from human induced pluripotent stem cells (hiPSCs)

Makorin ring finger protein 3 (MKRN3) was identified as an inhibitor of puberty initiation with the report of loss-of-function mutations in association with central precocious puberty. To investigate the roles and mechanisms of action of MKRN3 within the human hypothalamus, we used hypothalamic neurons derived from human induced pluripotent stem cells (hiPSCs). MKRN3 deletion was introduced into hiPSCs using CRISPR interference (CRISPRi) technology. Three separated hypothalamic differentiation of MKRN3-wildtype hiPSCs (MKRN3-WT I, II and III) and MKRN3-deficient hiPSCs (MKRN3-KO I, II and III) were performed using an established hypothalamic neuron differentiation protocol. To identify hypothalamic targets of MKRN3, we performed comparative transcriptome analysis by RNA sequencing (RNA-seq) of MKRN3-WT and MKRN3-KO hypothalamic neurons after 30 days of differentiation. Comparative gene expression profiling analysis of RNA-seq data for MKRN3-WT and MKRN3-KO hypothalamic neurons derived from hiPSCs. Three replicates issued from three different hypothalamic differentiations of MKRN3-wildtype hiPSCs (MKRN3-WT I, II and III) and MKRN3-deficient hiPSCs (MKRN3-KO I, II and III) were analyzed.