Our team aims to engage in a synthetic biology project that carries significant relevance to current day problems. Following a thorough project exploration phase, our team decided on a project titled, “Clean Green Lipid Machine” which involves the genetic engineering of microalgae.

Microalgae are a varied group of organisms with great potential in synthetic biology. Compared to other iGEM & genetic engineering model organisms, such as E. coli and yeast, microalgae distinguish themselves by the fact that they are photosynthetic with the potential of carbon neutral production of a variety of high valuable metabolites. These metabolites include biofuels, biolipids, proteins, hormones and antibodies, many of which are currently produced via resource intensive processes.

Our goal is to develop genetic engineering tools for microalgae, specifically various Chlorella & Chlamydomonas strains , to turn them into a chassis for future iGEM teams and researchers. This will provide an organism capable of synthesizing a variety of useful molecules such as lipids, which can be used as biofuels, or modified into useful lipid based compounds like omega 3 fatty acids.

One of our project goals is to create a library of useful and characterized parts; mainly by designing cassettes with varying promoters, genes and terminators from different species. These cassettes were constructed via Gibson assembly and then the plasmids containing the cassettes were inserted in Chlamydomonas and Chlorella via electroporation.

Our team grew out four different strains of Chlorella (Vulgaris, Ellipsodia, Saccharophilea, Kessleri) and a cell wall deficient strain of Chalmydomonas (UVM1). We characterized their growth patterns by obtaining various optical density and cell count measurements. With these results, we created growth curves and established the mid-log phase (ideal for transformation) of all of our strains.

We plan on using CrispR along with an NLS signal to direct the cas9 gene into the nucleus. By proving that our CrispR system works, we could safely assume that this system would work for the insertion of other genes of interest in Chlorella as well.