TY - THES A1 - Brons, Nico T1 - Implementation of a Genome Editing Procedure for Wheat (Triticum aestivium) Protoplasts with CRISPR-Cas9 Ribonucleoproteins N2 - Wheat (Triticum aestivium) is one of the most important crops in feeding the world. Even though its cultivation is subject to continuous optimization, there are still limitations to it. Wheat lodging, droughts and diseases are amongst the most prominent problems, which are only getting worse through climate change in the coming years. In 2012, a novel technique for fast and easy genetic modifications was discovered known as CRISPR-Cas9. By genetically modifying plants, it could be possible to alter them according to the needs of agriculture. Transgenic plants are usually obtained by infecting them with foreign DNA (plasmids), which usually results in a regulation by government. A novel DNA free approach described by (Brandt, Gunn et al. (2020)) might be the solution to this problem and was set as a guideline for the experiment conducted here. In this approach, wheat protoplasts were transfected with Cas9-Ribonucleoproteins (RNPs) to induce a deletion in their genome. After an incubation of 48 hours, the gene of interest was amplified by PCR and the resulting DNA fragments were analyzed by gel electrophoresis to detect successful deletions. Even though a detection of deletions was not successful, many limiting factors of the protoplasts’ treatment could be determined, and some could also be resolved. This experiment provides a great basis for future approaches. By using RNPs instead of DNA, no foreign DNA is integrated into the organism’s genome, which might result in a deregulation of plants altered with this technique. KW - Genome editing Y1 - 2022 ER -