The proto‐oncogene ets1 is highly expressed in the pre‐migratory and migratory neural crest (NC), and has been implicated in the delamination and migration of the NC cells. To identify the downstream target genes of Ets1 in this process, we did RNA sequencing (RNA‐Seq) on wild‐type and ets1 mutant X. tropicalis embryos. A list of genes with significantly differential expression was obtained by analyzing the RNA‐Seq data. We validated the RNA‐Seq data by quantitative PCR, and examined the expression pattern of the genes identified from this assay with whole mount in situ hybridization. A majority of the identified genes showed expression in migrating NC. Among them, the expression of microseminoprotein beta gene 3 (msmb3) was positively regulated by Ets1 in both X. laevis and X. tropicalis. Knockdown of msmb3 with antisense morpholino oligonucleotides or disruption of msmb3 by CRISPR/Cas9 both impaired the migratory streams of NC. Our study identified msmb3 as an Ets1 target gene and uncovered its function in maintaining neural crest migration pattern.

中文翻译：

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热带爪蟾ets1突变胚胎的RNA-Seq分析确定微精蛋白β基因3是神经嵴迁移的重要调节因子

原癌基因 ets1 在迁移前和迁移神经嵴 (NC) 中高度表达，并与 NC 细胞的分层和迁移有关。为了在此过程中鉴定 Ets1 的下游靶基因，我们对野生型和 ets1 突变体热带葡萄球菌胚胎进行了 RNA 测序（RNA-Seq）。通过分析 RNA-Seq 数据获得了具有显着差异表达的基因列表。我们通过定量 PCR 验证了 RNA-Seq 数据，并通过整体原位杂交检查了从该测定中鉴定的基因的表达模式。大多数鉴定的基因在迁移的 NC 中表现出表达。其中，微精蛋白β基因3（msmb3）的表达受Ets1在X. laevis和X.tropis中的正调控。用反义吗啉代寡核苷酸敲除 msmb3 或通过 CRISPR/Cas9 破坏 msmb3 都会损害 NC 的迁移流。我们的研究将 msmb3 鉴定为 Ets1 靶基因，并揭示了其在维持神经嵴迁移模式中的功能。