Cytoskeletal Function in C. elegans Embryos

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We request funds to purchase two high capacity and versatile BioRad Thermal Cyclers. These will replace a single much older and lower capacity thermal cycler currently shared by my lab and the adjacent labs of my nearby colleagues and faculty members Chris Doe and Diana Libuda. Currently, due to the age of our machine and its limited capacity, and a lack of other institutional options, we have suffered from lack of access to amplification for constructing transgenic strains we need for our research. The discovery of CRISPR/Cas9 and its applications to genome editing have revolutionized molecular genetics in all systems, including C. elegans, and our limited thermal cycler capacity is now frequently limiting and slowing our progress. By purchasing two new higher capacity machines, and having a shared arrangement with two other labs, we will relieve not only the limitation to access in my own lab but also in the labs of two of my closest and physically adjacent colleagues. The remarkable advances in high resolution light microscopy make our ability to generate fluorescent protein fusions to endogenous loci more and more powerful, and we also are implementing more highly mechanistic structure/function studies that require the creation of transgenic lines that express mutant transgenes to replace endogenous genes. Finally, we also are now beginning to use optogenetics to more rigorously explore gene requirements. We need increased thermal cycler capacity to generate transgene constructs for use in all of these powerful molecular genetic approaches.