Study 5- RNA-seq of male KOLF2.2J hiPSC-derived cortical brain organoids and extra-embryonic lineages homozygous null for nine different transcription factors and one RNA splicing factor

This study aimed to phenotype iPSC-derived cortical brain organoids that contain homozygous null alleles for transcription factors SIX3 and NR2F1 and to phenotype iPSC-derived extra-embryonic cell lineages that contain homozygous null alleles for transcription factors HEY1, HOPX, MAFF, TWIST2, DLX4, DLX5, and DLX6 and the splicing factor MBNL2. Null alleles were generated by the insertion of a premature termination codon with frameshift (PTC+1). This analysis provided insight into the impact of loss of function for these transcription factors in the respective cell lineages. The study involved the generation of RNA-seq on 48 samples, comprising 12 differentiated into cortical brain organoids (with dorsal forebrain patterning) and 36 differentiated into the trophoblast lineage, promoting the formation of a primitive syncytium. Three biological replicates were run for each gene KO line. Additionally, three biological replicates were run of wild-type KOLF2.2J cells (the parental male cell line in which the KOs were generated) under the same differentiation conditions.