CRISPR-Cas9 and CRISPR-Cpf1 Mediated Genome Editing of EPFL9 Gene in Rice

The development of the CRISPR-Cas9/Cpf1 genome editing system has provided a convenient tool for creating loss of function mutants for genes of interest. Xiaojia Yin led a team from the International Rice Research Institute in the Philippines and used CRISPR-Cas9 and CRISPR-Cpf1 technology to knock out the rice orthologue of the early developmental gene Epidermal Patterning Factor like-9 (EPFL9), a positive regulator of stomatal development in Arabidopsis thaliana.

Developed mutants showed edits in the target gene, which were inherited into the T2 generation. The mutant plants exhibited significant reduction in stomatal density on the lower leaf surface of the genome-edited rice plants. The team also used the CRISPR-LbCpf1 (Lachnospiracaebacterium Cpf1) to target the same OsEPFL9 and had almost identical results. There were also no significant off-target mutations detected.

This study demonstrated the application of CRISPR-Cas9/Cpf1 in developing heritable gene edits.

For more information, read the article in Plant Cell Reports.