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, Jessica; Greiner, Andre; Schellwat, Jana; Neukam, Martin; Rottmann, Theresa; Lu, Yinghong; Kateriya, Suneel; Hegemann, Peter & Kreimer, Georg
  
• (2013) Nuclear gene targeting in Chlamydomonas using engineered Zinc Finger Nucleases. Plant J. 73, 873 – 882
  Sizova, Irina; Greiner, Andre; Awasthi, Mayanka; Kateriya, Suneel & Hegemann, Peter
  
• (2016) A blue light photoreceptor mediates the feedback regulation of photosynthesis. Nature 537, 563 – 566
  Petroutsos, Dimitris; Tokutsu, Ryutaro; Maruyama, Shinichiro; Flori, Serena; Greiner, Andre; Magneschi, Leonardo; Cusant, Loic; Kottke, Tilman; Mittag, Maria; Hegemann, Peter; Finazzi, Giovanni & Minagawa, Jun
  
• (2017) An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis thaliana. Frontiers in Plant Science Vol. 8, Article 39
  Hahn, Florian; Mantegazza, Otho; Greiner, André; Hegemann, Peter; Eisenhut, Marion & Weber, Andreas P. M.