Gene Expression and Manipulation

项目总结-基因表达和操作 基因表达和操纵模块的重点是提供支持，协助和促进 与基因表达和操作有关的具体研究。我们支持的大多数研究类型都属于 眼分子生物学、生物化学和细胞生物学的范围。这包括定量评估， 治疗、条件或疾病状态对以下表达水平、mRNA和蛋白质的时间效应： 眼组织中的基因我们也支持研究集中在确定个别基因的影响， 操纵眼突和细胞。这包括敲低细胞中特定的基因表达水平 或组织中，通过RNAi沉默，通过过表达增加特异性基因表达，以及调节活性 或通过表达修饰形式，例如通过转染或感染遗传突变体， 显性失活、组成型活性或选择性剪接基因构建体。操纵方法包括 构建体转染和通过用重组病毒构建体感染的病毒基因或RNAi递送。表达 使用PCR阵列的研究和顺式元件启动子的研究也是容易的。我们最近一直在协助 CRISPR/Cas9基因编辑研究旨在建立疾病模型， 疾病相关的修饰和干细胞操作。我们拥有并维护的仪器支持 这些类型的研究。我们对所有这些方法都有经验，并将为核心用户提供便利和帮助 任何他们认为对调查有价值的东西除了向NEI资助的 调查员，我们积极支持调查员，特别是新的和年轻的调查员， 试点或初步数据，可以利用到未来的基金会和NEI资金。

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.