KeyBase®
How it works
KeyBase is a novel method for targeted mutagenesis of plant cells by creating deliberate point mutations in your gene of interest. It works through the introcution into plant protoplasts of mutagenic oligonucleotides, with sequence complementary to the target gene and containing chemically modified nucleotides to enhance the mutagenic process.
Benefits
The main benefit of KeyBase over the use of chemical mutagenesis is the power to create point mutations in predefined genomic positions in contrast to the random mutations obtained with conventional mutagenesis methods.
Applications
KeyBase can be used to make subtle changes in the genomic sequence of plants, that could result in: 1) modified amino acid sequences of proteins, 2) complete gene knockouts by introducing stopcodons or frameshift mutations, 3) modified gene expression by making changes in promoter sequences. Such mutations may be useful to inhibit unwanted gene expression, increase beneficial gene expression or change proteins and enzymes in more stress-tolerant or enzymatically active variants.
Deliverables
A prerequisite of applying KeyBase is a fairly efficient protoplast system capable of regenerating whole plants. Such protoplasts systems are available for many crop plants. Deliverables of a KeyBase project are healthy growing plants in which the desired pointmutations have been confirmed by sequencing.
Examples
Examples of KeyBase mutants that have already been created are herbicide-tolerant tobacco, tomato and Petunia plants, e.g. plants that resist commercial applications of sulfonylurea herbicides. With the further development of KeyBase into a more efficient method, non-selectable targets will become possible, such as mutants with increased abiotic stress tolerance, increased insect or virus resistance, increased yield and altered flower colors in ornamentals.