(5, 6) It is the combination of these two principles-predictable parts and rapid prototyping-that give synthetic biologists the ability to approach difficult problems in energy, (7, 8) agriculture, (9) and human health. (1-4) Because manipulations to the cell often lead to unexpected results, progress is best made by rapidly iterating through highly parallelized experiments to explore a wide parameter space. These challenges have led to an additional, and equally important, aspect to synthetic biology: rapid prototyping. Despite these efforts, the inner workings of the cell continue to elude understanding, and while certain elements can be highly predictable, the system behavior as a whole is difficult to anticipate. Synthetic biologists have sought ways to abstract the layers of complexity into components with predictable interactions, making it more feasible to undertake large engineering projects. Unfortunately, the potential power of the cell also brings with it a level of complexity that makes engineering biological systems extremely difficult. Synthetic biology is driven by the desire to engineer novel biological functions that push the boundaries of what can be accomplished within living cells. Additionally, by relieving researchers of the burden of technical details, they can focus on higher-level aspects of experimental design. With this toolkit, we strive to simplify the process of engineering yeast by standardizing the physical manipulations and suggesting best practices that together will enable more straightforward translation of materials and data from one group to another. Additionally, we describe genome-editing tools for making modifications directly to the yeast chromosomes, which we find preferable to plasmids due to reduced variability in expression. This platform provides a framework in which to create new designs, as well as data on promoters, terminators, degradation tags, and copy number to inform those designs. Here we present a versatile engineering platform for yeast, which contains both a rapid, modular assembly method and a basic set of characterized parts. While there is a wealth of resources and literature about the biology of yeast, it can be daunting to navigate and extract the tools needed for engineering applications. However, until recently, most synthetic biology systems have focused on bacteria. Saccharomyces cerevisiae is an increasingly attractive host for synthetic biology because of its long history in industrial fermentations.

Slic toolkit