General guidelines for CRISPR/Cas-based genome editing in plants.

Area of Science:

• Agricultural Science
• Molecular Biology
• Biotechnology

Background:

• CRISPR/Cas9 is a precise genome editing tool for enhancing plant traits.
• It offers advantages over conventional breeding and standard genetic engineering for developing climate-resilient crops.

Purpose of the Study:

• To review CRISPR/Cas9 components and methods for plant genome editing.
• To provide guidelines for developing genome-edited plants from sgRNA design to T2 generation selection.

Main Methods:

• Utilizing bioinformatics tools for single guide RNA (sgRNA) design and off-target prediction.
• Employing various vectors, promoters, Cas proteins, and terminators for gene editing.
• Applying tissue culture-based plant transformation techniques.

Main Results:

• CRISPR/Cas9 enables precise modification of plant genomes for trait improvement.
• Multiple-target sgRNA designs allow simultaneous silencing of gene families or pathways.
• Guidelines are provided for efficient genome editing workflows.

Conclusions:

• CRISPR/Cas9 technology is crucial for rapid development of climate-resilient plant varieties.
• Effective sgRNA design and understanding of CRISPR/Cas9 components are essential for successful genome editing.
• This review offers a comprehensive guide for researchers in plant genome editing.