More than 99% of all genes of C. reinhardtii are encoded in the nuclear DNA. Over the past decades, classical genetic techniques and molecular biological strategies have been established that allow manipulation of the nuclear DNA for fundamental research and applied tasks. The nuclear genome is haploid during the vegetative stage of the green microalga’s life cycle, a feature promoting the elucidation of specific genotype–phenotype interrelations in genetic studies. Supported by the general progress of molecular technologies, the toolbox for C. reinhardtii nuclear genome transformation and engineering has been greatly expanded, and, today, a substantial arsenal of tools and techniques is available which can... More
More than 99% of all genes of C. reinhardtii are encoded in the nuclear DNA. Over the past decades, classical genetic techniques and molecular biological strategies have been established that allow manipulation of the nuclear DNA for fundamental research and applied tasks. The nuclear genome is haploid during the vegetative stage of the green microalga’s life cycle, a feature promoting the elucidation of specific genotype–phenotype interrelations in genetic studies. Supported by the general progress of molecular technologies, the toolbox for C. reinhardtii nuclear genome transformation and engineering has been greatly expanded, and, today, a substantial arsenal of tools and techniques is available which can be utilized to approach individual molecular research tasks.
Several important aspects of nuclear genetic engineering with C. reinhardtii will be addressed in this chapter. Since in many cases, the prerequisite for genetic engineering is the transformation of DNA, strategies for nuclear DNA delivery will be introduced within the historical context. Furthermore, established DNA elements, transformation selection markers, strategies to achieve modulation of gene expression, and examples of versatile vectors will be summarized. Finally, cell line-specific considerations will be addressed before future perspectives are presented.
