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
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