Investigating retinal development through loss of function analysis

During retinal development, six different general classes of neurons are produced. Of these six classes of neurons, it is not well understood how a variety of genetic factors work together to control the differentiation of these nerve cells. Previously, there have been genes identified in a large-scale screen of all the mRNAs expressed in individual developing retinal cells. The goal of my project is to design an RNA interference (RNAi) based strategy to alter the expression of particular genes during mouse retinal development. Specifically, my objective has been to design and test RNAi constructs designed to five different genes. To accomplish this goal I have set up a tissue culture based sensor system with portions of each mRNA linked to GFP. Therefore, when a successful RNAi construct is introduced into cells, the GFP expression is significantly knocked down. Constructs that work well in these experiments are then introduced into the developing mouse retina. Any resulting phenotypes will be observed using a combination of immunostaining and qPCR. Through these knockdown experiments we will learn more about how specific genetic factors work to control the development of different retinal neuron