Original title:
Insertion of CRISPR system to carrot to enable genome editing
Authors:
Chaloupský, Pavel Document type: Master’s theses
Year:
2018
Language:
eng Abstract:
Main goal of this study was to develop carrot plants and callus lines with stable expression of Cas9 and Cpf1 genes. To achieve the thesis objective, transformation of Escherichia coli and Agrobacterium tumefaciens strains with different codon-optimized CRISPR constructs was carried out. Agrobacterium-mediated carrot callus transformation with CRISPR constructs, induction of somatic embryogenesis, and molecular confirmation of transgenic events in carrot were also part of the experiment executed within this thesis. By use of PCR-based genotyping, transgene was confirmed in 346 calli. Relative expression of Cas9 and Cpf1 in 15 selected lines was measured.
Keywords:
Agrobacterium; Cas9; Cpf1; Daucus carota; somatická embrzogeneze