My goal in the current projects is to find ways for algae to accumulate more neutral lipids. Although genomic, transcriptomic and proteomic analyses of algae for biofuel production have been widely used, these experiments did not find simple targets for improvement by bioengineering. We are trying to overcome these complex problems in various ways. First, we found several enzymes that seem to be uniquely involved in TAG synthesis in Chlamydomonas. One of them is CrAGPAT, an acyltransferase that prefers C16 over C18 fatty acids, although it is localized outside of the chloroplast. We are also trying to overexpress multiple enzymes/transporters at the same time in Chlamydomonas, to change algal metabolism towards neutral lipid synthesis. We have already targeted BASS and a pyruvate kinase (see below). Finally, we are interested in the transcriptional regulation that occurs under nutritional starvation in algae. We have chosen promoter regions that potentially give strong gene expression under nitrogen starvation conditions. The promoters will be fused with reporter genes, and reporter strains will be isolated and utilized to identify mutants with altered reporter expression.