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Fabian-Philipp Sylvester (FU Berlin)

Fabian-Philipp Sylvester's Master thesis was awarded with the Prize for the Best Plant Science Master Thesis, which was carried out at FU Berlin in the year 2018.

Title of awarded thesis

"Targeted „genome editing“ via CRISPR/Cas9 and characterization of CDPKs in stress response"

This work shows the great potential of CRISPR/Cas9 regarding the analysis of coupled inherited isoforms of a gen family, here a CDPK gene cluster. Sylvester for the first time generated a triple mutant including cpk21, cpk23 using CRISPR/Cas9.

In the literature homologous calcium-dependent protein kinases (CDPK also CPK) CPK21 and CPK23 from Arabidopsis thaliana share similar function in the phytohormone abscisic acid-dependent stress response. This data suggest an at least partial functional redundancy. The generation of a double mutant line by crossing is prevented through the adjacent localization of both genes coding for CPK21 and CPK23 on chromosome 4. Therefore, in this thesis a cpk21cpk23 double mutant was generated via targeted genome editing. The resulting double mutant and respective single mutants were compared and assessed in germination rate, root length, biomass and rosette diameter. These phenotypical analyses demonstrated a reduced root length of 5-day old seedlings in cpk21 and cpk23 mutants, which was not enhanced in the double mutant. To investigate a potential contribution of a different isoform, in this master thesis a triple mutant including cpk21, cpk23 was generated for the first time via CRISPR/Cas9.

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Fabian-Philipp Sylvester conducted this work at the department biochemistry of plants in the group of Prof. Dr. Tina Romeis