FUNCTIONAL GENOMICS

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心机构，不一定为研究者机构。功能基因组学模块的三个组成部分是：1）用于测试转基因小鼠基因功能的设施。我们创建了一个最先进的小鼠遗传学设施，主要支持通过小鼠胚胎干细胞的靶向诱变来操纵小鼠基因组。该设施使我们能够通过对小鼠胚胎干细胞进行精确的基因组修饰来生成人类疾病和病理的动物模型。2)在细胞培养中使用RNA干扰进行反向遗传学的体外设施。RNAi是一种引入序列特异性双链小抑制RNA（siRNA）的技术，已成为一种在分离细胞中敲低基因表达的有力工具。siRNA可以被引入到在化学定义的环境中生长的视网膜神经元中，以研究许多基因的功能。可以在不受整个生物体的稳态影响的情况下评价使用体外系统对内源性和转染基因表达的抑制。3)在动物模型中使用RNA干扰进行反向遗传学的体内设施。通过siRNA分子在完整视网膜的终末分化细胞中的长期表达来研究iRNA的治疗潜力。我们成功地实施了这个功能基因组学模块，缩短了我们的COBRE资助的PJIs提交首次R 01申请的时间，并扩大了我们营地的其他视觉研究人员，模块主任和/或COBRE导师的研究视野

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The three components of the Functional Genomics Module are: 1) A facility that will be used to test gene function in genetically modified mice. We created a state-of-the-art mouse genetics facility that will mainly support the manipulation of the mouse genome by targeted mutagenesis in mouse embryonic stem cells. This facility allows us to generate animal models of human diseases and pathology through precise genomic modifications in mouse embryonic stem cells. 2) An in vitro facility for reverse genetics using RNA interference in cell culture. RNAi, the introduction of sequence-specific double-stranded small inhibiting RNAs (siRNAs), has become a powerful tool to knock down gene expression in isolated cells. siRNAs can be introduced into retinal neurons grown in a chemically defined environment to study the function of numerous genes. The inhibition of the expression of endogenous and transfected genes using in vitro systems can be evaluated free from the homeostatic influence of the whole organism. 3) An in vivo facility for reverse genetics using RNA interference in animal models. The therapeutic potential of iRNA is investigated by long-term expression of the siRNA molecules in the terminally differentiated cells of the intact retina. Our successful implementation of this Functional Genomics Module has shortened the time for submission of first-time R01 applications by our COBRE-funded PJIs and expanded the research horizons for other vision researchers, module directors, and/or COBRE mentors on our camp