Abstract Until recently, precise genome editing has been limited to a few organisms. The ability of Cas9 to generate double stranded DNA breaks at specific genomic sites has greatly expanded molecular toolkits in many organisms and cell types. Before CRISPR‐Cas9 mediated genome editing,P. patenswas unique among plants in its ability to integrate DNA via homologous recombination. However, selection for homologous recombination events was required to obtain edited plants, limiting the types of editing that were possible. Now with CRISPR‐Cas9, molecular manipulations inP. patenshave greatly expanded. This protocol describes a method to generate a variety of different genome edits. The protocol describes a streamlined method to generate the Cas9/sgRNA expression constructs, design homology templates, transform, and quickly genotype plants. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Constructing the Cas9/sgRNA transient expression vector Alternate Protocol 1: Shortcut to generating single and pooled Cas9/sgRNA expression vectors Basic Protocol 2: Designing the oligonucleotide‐based homology‐directed repair (HDR) template Alternate Protocol 2: Designing the plasmid‐based HDR template Basic Protocol 3: Inducing genome editing by transforming CRISPR vector intoP. patensprotoplasts Basic Protocol 4: Identifying edited plants. 
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                            Heterozygous fasciated ear mutations improve yield traits in inbred and hybrid maize lines
                        
                    
    
            Heterozygous mutations in two genes encoding key regulators of development improve kernel row number in inbred and hybrid maize, revealing their potential for yield improvement. 
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                            - Award ID(s):
- 2129189
- PAR ID:
- 10627424
- Publisher / Repository:
- American Society of Plant Biologists (ASPB)
- Date Published:
- Journal Name:
- Plant Physiology
- Volume:
- 196
- Issue:
- 4
- ISSN:
- 0032-0889
- Page Range / eLocation ID:
- 2291 to 2295
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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