Recent progress in plant genome engineering: from large insertions to chromosome number changes.

Area of Science:

• Biotechnology
• Molecular Biology
• Genomics

Background:

• The CRISPR/Cas system is a versatile tool for targeted genome modification.
• Previous genome editing methods were limited to small sequence alterations.

Purpose of the Study:

• To review advances in CRISPR/Cas-based genome engineering for large-scale modifications.
• To highlight new opportunities in crop improvement and synthetic biology.

Main Methods:

• Homology-directed gene targeting for kilobase insertions.
• CRISPR/Cas with recombinases/transposases for larger sequence integration.
• Prime editing for megabase-scale DNA alterations.
• Chromosome fission and fusion for karyotype engineering.

Main Results:

• Efficient, scar-free kilobase insertions are now possible.
• Integration of large DNA sequences is achievable.
• Prime editing enables extensive DNA modifications (hundreds of kilobases to megabases).
• Controlled chromosome restructuring through fission and fusion has been demonstrated.

Conclusions:

• Genome engineering has advanced to enable large-scale modifications and karyotype restructuring.
• These techniques offer novel strategies for crop improvement, including trait stacking on Plant Artificial Chromosomes.
• The ability to mimic evolutionary processes and establish reproductive barriers is now feasible.