Calysta’s proprietary biological GTL platform utilizes genetically engineered methanotrophs - prokaryotes that utilize methane as their sole source of carbon and energy. Methanotrophs have been observed in a wide range of environments, both aerobic and anaerobic, typically in association with natural methane sources such as degrading biomass or petroleum offgas. While methanotrophs are a logical starting point for the development of a biological methane conversion platform, a critical requirement for the development of a biotechnology platform is the availability of tools for the directed manipulation and modification of the host cell’s metabolism. Although such tools are commonplace for model organisms (e.g. Escherichia coli or Saccharomyces cerevisiae), relatively little effort has been expended to develop similar capabilities in methanotrophs. Calysta has therefore developed a suite of tools for the expression of heterologous proteins in methanotrophs, as well as tools for the efficient targeted manipulation of the methanotroph genome. This presentation will focus on our recent advances and developments to this toolbox.