Generating transgenic fluorescent C. elegans for tissue-specific gene expression profiling

Studying the genetic contribution to a complicated disease in a tissue-specific context requires a comprehensive view of gene expression in each affected tissue of an organism. Current approaches in creating such a tissue- or cell-specific gene expression profile are limited. The common approaches, such as single molecule in situ hybridization (smFISH), provide temporal and spatial expression profiles for target genes; however, this method is costly and low-throughput due to the necessity for probe designing and limited fluorescent channels. The high throughput methods, such as RNA sequencing, require sample homogenization, and therefore, all spatial information is lost. Our lab is developing a new high-throughput technique to acquire a tissue-specific gene expression profile using the animal model Caenorhabditis elegans. One of the bottlenecks for developing this technique is the visualization of cell boundaries. Fluorescent constructs labeling cell membranes were generated by previous studies; however, all of the currently available constructs are tissue-restricted. Therefore, it is necessary to create new generic fluorescent constructs enabling expression of a cell boundary marker in all cells. This study focuses on generating two new DNA constructs by plasmid cloning methods. The plasmid vectors containing these constructs will be further validated and injected into C. elegans to generate transgenic fluorescent strains for tissue-specific gene expression profiling.