Creation and manipulation of bipartite expression transgenes inC. elegansusing phiC31 recombinase

Abstract

AbstractBipartite expression systems are widely used in model organisms to express specific gene products in a cell-specific context. They are typically encoded as two independent, unlinked transgenic insertions: a driver and a reporter. Herein, I outline an efficient method namedrecombination-mediated homologexchange (RMHE) that uses genetically encoded phiC31 recombinase to convert driver and reporter transgenes integrated at the same genetic position from atransconfiguration where one component is on each chromosome, to acisconfiguration where the driver and reporter are on the same chromosome. This approach is based upon the development of a set of 3’attBtagged driver lines and 5’attPtagged reporter lines. This genetic based approach leverages both the power of combinatorial re-assortment of drivers and reporters and the simplicity of single locus genetics. I also describe a novel microinjection-based approach namedrecombination-mediated integration (RMI) that utilizes the individualattBdriver andattPreporter lines as landing site for the phiC31 recombinase mediated integration of whole driver and reporter plasmids into the existing reporter and driver lines, respectively. Thus, this work outlines both a novel genetic based and a novel microinjection-based method to create cis-linked driver/reporter pairs. These new tools increase the utility of bipartite systems forC. elegansgenetics by reducing the complexity of reporter system segregation in crosses and thus can greatly simplify the use of bipartite reporter systems during genetic analysis.