Central Project

• Applicants: Professor Dr. Peter Hegemann; Professorin Dr. Maria Mittag; Professor Dr. Andreas P.M. Weber
• Subject Area: Plant Physiology
• Term: from 2013 to 2016
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 112805507

• Final Report Year: 2017

Final Report Abstract

Within the Z-project of FOR1261 we have established three different gene-editing strategies and provided protocols for rapid isolation of Chlamydomonas mutants from wild-type-like strains. Genetically encoded zinc-finger nucleases or Cas9 from Staphylococcus aureus and Streptococcus pyogenes as well as recombinant Cas9 protein inactivated non-selectable genes e.g. for phytoene synthase-1, channelrhdopsin-1 and 2, chlamyopsin-1, a voltage gated Ca-channel, phototropin, the UV-receptor UVR8, and a histidin-kinase-rhodopsin-1. After careful adaptation of transformation protocols and donor DNA design 5-15% of analyzed clones contained inactivated genes of interest.

Publications

• (2012) Phototropin, a Novel Player in Eyespot Development and Control of Phototactic Behaviour in Chlamydomonas reinhardtii. Plant Cell 24:4687-702
  Trippens, J., Greiner, A., Schellwat, J., Neukam, M., Rottmann, T., Lu, Y., Suneel Kateriya, S., Hegemann, P., and Kreimer, G.
  (See online at https://doi.org/10.1105/tpc.112.103523)
• (2013) Nuclear gene targeting in Chlamydomonas using engineered Zinc Finger Nucleases. Plant J. 73, 873 – 882
  Sizova, S., Greiner, A., Awasthi, M., Kateriya, S., and Hegemann, P.
  (See online at https://doi.org/10.1111/tpj.12066)
• (2016) A blue light photoreceptor mediates the feedback regulation of photosynthesis. Nature 537, 563 – 566
  Petroutsos, D., Tokutsu, R., Maruyama, S., Flori, S., Greiner, A., Magneschi, L., Cusant, L., Kottke, T., Mittag, M., Hegemann, P., Finazzi, G., Minagawa, J.
  (See online at https://doi.org/10.1038/nature19358)
• (2017) An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis thaliana. Frontiers in Plant Science Vol. 8, Article 39
  Hahn, F., Mantegazza, O., Greiner, A., Hegemann, P., Eisenhut, M. and Weber, A.P.M.
  (See online at https://doi.org/10.3389/fpls.2017.00039)