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

Rubisco是在包括高等植物在内的绝大多数光合生物中负责CO2同化的酶。它的效率相对较低，数十年的研究一直试图提高其对CO2的亲和力和特异性。成功是非常有限的，在很大程度上是因为缺乏工具来操纵植物中的Rubisco特性（1）。越来越多的基于合成生物学的工具现在使植物基因组能够以前所未有的精度进行编辑。该项目的目的是开发一种方法，用选择的异源核编码小亚基（SSU）取代拟南芥中Rubisco酶的核编码小亚基（SSU）家族。这种方法将提供前所未有的能力来评估Rubisco SSU特性在一系列条件下确定光合效率的作用（2）。它开辟了前景，提高作物产量在未来通过一种新的遗传剪裁战略。培训：该项目是一个优秀的培训机会，发展多学科的方法，植物科学。您将在广泛的分子和整个植物分析技能方面发展专业知识，包括DNA，RNA和蛋白质分析（例如qRT-PCR，Western blot，酶活性测定），以及使用叶片气体交换和荧光技术的光合生理学。您将通过使用CRISPR/cas9系统与转基因拟南芥品系合作，获得基于合成生物学的方法的经验（3）。这些品系将用于直接和明确地测试天然SSU的取代是否可以导致i）功能性Rubisco全酶的组装，ii）改善Rubisco性能，iii）增加植物生长速率。本研究为Rubisco在植物中的调控和组成以及SSU在植物生长发育中的作用等基础和应用研究提供了重要的新信息。您将与作物科学家互动，制定策略，将重要发现应用于作物。您还将有大量机会在定期会议上展示您的研究，包括国家和国际会议。

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.