Gene Expression and Manipulation Core

PROJECT SUMMARY - GENE EXPRESSION & MANIPULATION The Gene Expression & Manipulation core is focused on providing support and assisting and facilitating specific studies relating to gene expression and manipulation. Most of the types of studies supported by this core 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 gene expression and protein levels 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 using vectors, 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, gene or RNAi delivery via infection with recombinant viral constructs such as lentivirus, or by lipid nanoparticles. We have recently been assisting investigators with CRISPR/Cas9 gene editing, including base and prime editing studies aimed at disease modeling with the intention of future disease related modifications or stem cell manipulation. This core maintains various instruments that support these types of studies. The core co-Directors and personnel have experience with all of these methods and will facilitate and assist core users with any of the described techniques that they deem of value to their investigations. In addition to providing this service to NEI-funded investigators, we actively support efforts of new and young investigators, to develop pilot or preliminary data that can be leveraged into future foundation and NEI funding.