Re-engineering photosynthesis: targeted genome editing to replace nuclear-encoded Rubisco in higher plants

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Rubisco is the enzyme responsible for CO2 assimilation in the vast majority of photosynthetic organisms, including higher plants. It is relatively inefficient, and decades of research have gone into attempting to improve its affinity and specificity for CO2. Success has been very limited, in large measure because of a dearth of tools with which to manipulate Rubisco properties in planta (1).A growing set of synthetic biology-based tools now enable plant genomes to be edited with unprecedented precision. The aim of this project is to develop a means of replacing the family of nuclear-encoded small subunits (SSUs) of the Rubisco enzyme in Arabidopsis with heterologous SSUs of choice. This approach will provide an unprecedented capacity to evaluate the role of Rubisco SSU properties in determining photosynthetic efficiency in a range of conditions (2). It opens up the prospect of enhancing crop yields in the future through a novel genetic tailoring strategy.Training: The project is an outstanding training opportunity for developing a multidisciplinary approach to plant science. You will develop expertise in a wide range of molecular and whole plant analysis skills, including DNA, RNA and protein analyses (e.g. qRT-PCR, Western blot, enzyme activity assays), and photosynthetic physiology using leaf gas exchange and fluorescence techniques. You will gain experience in synthetic biology-based approaches by working with transgenic Arabidopsis lines using the CRISPR/cas9 system (3).These lines will be used to directly and unambiguously test if substitution of native SSUs can lead to i) the assembly of functional Rubisco holoenzymes, ii) improved Rubisco performance, and iii) increased plant growth rates. This work will provide important new information for both fundamental and applied research about the regulation and makeup of Rubisco in plants and the role of the SSU in growth and development. You will interact with crop scientists to develop strategies to take important findings through to application in crops. There will also abundant opportunity for you to present your research at regular meetings, including national and international conferences.