Role of KLF4 and KLF5 in mouse cornea development

描述（由申请人提供）：本研究计划旨在了解Kruppel样锌指转录因子KLF4和KLF5（小鼠角膜中最丰富的两种转录因子）在调节成熟角膜正常发育和维持中的作用。KLF4和KLF5基因敲除小鼠的早期致死性是研究其参与调节角膜正常发育的障碍，角膜正常发育持续至出生后6周。为了克服这个问题，本文提出了KLF4和KLF5基因的条件性破坏。将通过组织学研究这些条件突变对角膜胚胎发育和角膜上皮细胞出生后分层的影响。将来自这些条件突变体的成熟角膜与野生型同窝仔的成熟角膜在透明度、形态学和伤口愈合能力方面进行比较。通过微阵列将它们的基因表达模式与野生型小鼠进行比较，并通过定量PCR和原位杂交证实差异。将在培养的细胞中分析KLF4和KLF5参与调节鉴定的感兴趣的靶基因的启动子。本文的研究将为角膜正常发育和维持过程中基因表达的调控提供重要的信息，并为进一步研究角膜基因调控这一相对不足的领域提供坚实的基础。具体目标：1。目的研究条件性破坏小鼠眼KLF4和KLF5基因对成熟角膜发育和维持的影响。2.目的：鉴定角膜发育过程中KLF4和KLF5的靶基因。3.研究KLF4和KLF5参与角膜中选定基因表达的调节。

DESCRIPTION (provided by applicant): This research proposal aims at understanding the role of Kruppel-like zinc-finger transcription factors KLF4 and KLF5, two of the most abundant transcription factors in the mouse cornea, in regulating the normal development and maintenance of mature cornea. Early lethality of KLF4 and KLF5 knockout mice is an impediment to study their involvement in regulating the normal development of cornea, which continues up to 6 weeks after birth. Conditional disruption of KLF4 and KLF5 genes is proposed here to overcome this problem. Effects of these conditional mutations on embryonic development of cornea and post-natal stratification of corneal epithelial cells will be studied by histology. Mature cornea from these conditional mutants will be compared with that of wild type littermates, with respect to transparency, morphology, and wound healing ability. Their gene expression patterns will be compared with that of wild type mice by microarrays and differences confirmed by quantitative PCR and in situ hybridizations. The involvement of KLF4 and KLF5 in regulating the promoters of identified target genes of interest will be analyzed in cultured cells. Studies proposed here will provide information of fundamental importance on regulation of gene expression during normal development and maintenance of cornea, and provide a solid foundation for further research in this relatively understudied field of gene regulation in cornea. Specific Aims: 1. To study the effect of conditional disruption of KLF4 and KLF5 genes in the mouse eye on the development and maintenance of mature cornea. 2. To identify the target genes for KLF4 and KLF5 in the developing cornea. 3. To study the involvement of KLF4 and KLF5 in regulation of expression of selected genes in the cornea.