Targets of Sf3b4 during neural crest development

Mutations in Sf3b4, a component of the spliceosome, have been linked to craniofacial dismorphology known as Nager syndrome. To understand the role Sf3b4 in this condition, we knockdown Sf3b4 in Xenopus laevis embryos, and found that in these embryos the formation of neural crest-derived craniofacial skeletal structures was compromised. We performed RNA-Seq to identify the repertoire of genes affected by Sf3b4 splicing activity. Overall design: Xenopus laevis embryos were injected at the 2-cell stage with mRNAs encoding noggin and wnt8 to induce neural crest progenitors. A subset of these embryos were co-injected with Sf3b4MO antisense oligonucleotides to interfere with Sf3b4 fonction during neural crest forrmation. At the blastula stage (stage 8/9), animal cap explants were dissected and cultured for 8 hours in a defined medium. Either two or three biological replicates were performed for each condition.