Gene Expression and Manipulation

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PROJECT SUMMARY - GENE EXPRESSION AND MANIPULATION The Gene Expression and Manipulation Module is focused on providing support, and assisting and facilitating specific studies relating to gene expression and manipulation. Most of the types of studies we support fall within the range of ocular molecular biology, biochemistry and cell biology. This includes evaluating, quantitatively, the temporal effects of treatments, conditions or disease states on the expression levels, mRNA and protein, of genes in ocular tissues. We also support studies focused on determining the effects of individual gene manipulation on ocular processes and cells. This includes knockdown of specific gene expression levels in cells or tissues by RNAi silencing, increases in specific gene expression by overexpression, and modulation of activity or function of specific genes by expressing modified forms, e.g. by transfecting or infecting genetic mutant, dominant negative, constitutively active or alternatively spliced gene constructs. Manipulation methods include construct transfection and viral gene or RNAi delivery via infection with recombinant viral constructs. Expression studies using PCR Arrays and cis element promoter studies are also facilitated. We have recently been assisting investigators with CRISPR/Cas9 gene editing studies aimed at disease modeling with the intention of future disease related modifications and stem cell manipulation. We have and maintain instrumentation that supports these types of studies. We have experience with all of these methods and will facilitate and assist core users with any that they deem of value to their investigations. In addition to providing this service to NEI-funded investigators, we actively support efforts of investigators, particularly new and young investigators, to develop pilot or preliminary data that can be leveraged into future foundation and NEI funding.