CRISPR-induced indels and base editing using the Staphylococcus aureus Cas9 in potato

Area of Science:

• Plant Science
• Genetics
• Biotechnology

Background:

• Genome editing, particularly CRISPR technology, is crucial for plant science, enabling precise genetic modifications.
• CRISPR systems effectively create knock-out mutants but have limitations like PAM restriction.
• Emerging CRISPR tools like base and prime editing offer nucleotide conversion for fine-tuning protein function.

Purpose of the Study:

• To demonstrate the efficacy of the Staphylococcus aureus Cas9 (SaCas9) system for genome editing in cultivated potato (Solanum tuberosum).
• To develop and apply a novel S. aureus-cytosine base editor for precise nucleotide conversions in potato.
• To expand the CRISPR toolbox for dicotyledonous plants, enhancing precision breeding applications.

Main Methods:

• Utilized the CRISPR-Cas9 system from Staphylococcus aureus (SaCas9) for inducing frameshift mutations.
• Developed a S. aureus-cytosine base editor to achieve targeted nucleotide conversions in the potato genome.
• Applied these tools to the tetraploid genome of Solanum tuberosum.

Main Results:

• Successfully introduced frameshift mutations in the tetraploid potato genome using SaCas9.
• Demonstrated the capability of the developed S. aureus-cytosine base editor for precise genetic modifications.
• Established proof-of-concept for advanced CRISPR applications in potato.

Conclusions:

• The SaCas9 system and the novel base editor are effective tools for genome editing in potato.
• These advancements broaden the CRISPR toolbox for dicotyledonous plants.
• The findings support enhanced biotechnology and precision breeding applications in cultivated potato.