Abstract：
Craniofacial development is controlled by a large number of genes, which interact with one another to form a complex gene regulatory network (GRN). Key components of GRN are signaling molecules and transcription factors. Therefore, identifying targets of core transcription factors is an important part of the overall efforts toward building a comprehensive and accurate model of GRN. LHX6 and LHX8 are transcription factors expressed in the oral mesenchyme of the first pharyngeal arch (PA1), and they are crucial regulators of palate and tooth development. Previously, we performed genome-wide transcriptional profiling and chromatin immunoprecipitation to identify target genes of LHX6 and LHX8 in PA1, and described a set of genes repressed by LHX. However, there has not been any discussion of the genes positively regulated by LHX6 and LHX8. In this paper, we revisited the above datasets to identify candidate positive targets of LHX in PA1. Focusing on those with known connections to craniofacial development, we performed RNA in situ hybridization to confirm the changes in expression in Lhx6;Lhx8 mutant. We also confirmed the binding of LHX6 to several putative enhancers near the candidate target genes. Together, we have uncovered novel connections between Lhx and other important regulators of craniofacial development, including Eya1, Barx1, Rspo2, Rspo3, and Wnt11.
摘要：
颅面发育受大量基因控制,它们相互作用形成复杂的基因调控网络(GRN)。GRN的关键组分是信号分子和转录因子。因此,识别核心转录因子的靶标是建立全面，准确的GRN模型的整体努力的重要组成部分。LHX6和LHX8是在第一咽弓（PA1）的口腔间质中表达的转录因子，它们是腭和牙齿发育的关键调节剂。以前,我们进行了全基因组转录谱分析和染色质免疫沉淀,以鉴定PA1中LHX6和LHX8的靶基因,并描述了一组被LHX抑制的基因.然而,还没有任何关于LHX6和LHX8正调控基因的讨论。在本文中,我们重新访问了上述数据集以确定PA1中LHX的候选阳性靶标.专注于那些与颅面发育有已知联系的人，我们进行RNA原位杂交以确认Lhx6;Lhx8突变体中表达的变化。我们还证实了LHX6与候选靶基因附近的几种推定增强子的结合。一起,我们发现了Lhx和其他重要的颅面发育调节因子之间的新联系，包括Eya1、Barx1、Rspo2、Rspo3和Wnt11。
