Genome editing in maize directed by CRISPR-Cas9 ribonucleoprotein complexes

Area of Science:

• Plant biotechnology
• Molecular biology
• Genetics

Background:

• Targeted DNA double-strand breaks enhance genome editing efficiency and precision.
• CRISPR-Cas9 is a versatile and widely adopted genome editing technology.
• Current plant genome editing often relies on DNA vector delivery of reagents.

Purpose of the Study:

• To report a novel method for delivering CRISPR-Cas9 ribonucleoproteins into maize embryo cells.
• To demonstrate DNA- and selectable marker-free gene mutagenesis in maize.
• To regenerate plants with edited alleles using this new delivery system.

Main Methods:

• Biolistic delivery of pre-assembled Cas9-gRNA ribonucleoproteins into maize embryo cells.
• Regeneration of edited maize plants from treated embryo cells.
• Assessment of gene mutagenesis and allele editing frequencies.

Main Results:

• Successful regeneration of maize plants with mutated and edited alleles.
• High-frequency DNA- and selectable marker-free gene mutagenesis achieved.
• Demonstration of efficient genome editing in maize using ribonucleoprotein delivery.

Conclusions:

• Biolistic delivery of ribonucleoproteins offers an efficient, DNA-free method for maize genome editing.
• This approach accelerates crop improvement and breeding practices.
• The method holds potential for application across diverse crop species.