Targeted Mutagenesis in Cotton using CRISPR-Cas9

The CRISPR-Cas9 system has been widely used for genome editing in various plants because of its simplicity, high efficiency and design flexibility. However, there are only a few reports on the application of CRISPR-Cas9-mediated targeted mutagenesis in cotton.

The team led by Xiugui Chen from the Chinese Academy of Agricultural Sciences and Huazhong Agricultural University reports genome editing in upland cotton (Gossypium hirsutum) using the CRISPR/Cas9 system. The team designed two guide RNAs to target distinct sites of the cotton Cloroplastos alterados 1 (GhCLA1) and vacuolar H⁺-pyrophosphatase (GhVP) genes. Mutations in these two genes were detected in cotton protoplasts.

The vectors were then transformed into cotton shoot apexes through Agrobacterium-mediated transformation, resulting in efficient target gene editing. Most of the mutations were nucleotide deletions, and the mutation efficiencies were 47.6–81.8% in genome-edited cotton plants. No off-target mutations were also detected.

These results indicate that the CRISPR-Cas9 system is an efficient and specific tool for targeted mutagenesis of the cotton genome.

For more information, read the article in Scientific Reports.