Study Finds Cas12a More Effective than Cas9 in Generating Knockout Mutants in MiRNA Genes in Rice

A study published in the Plant Biotechnology Journal investigates the effectiveness of the CRISPR-Cas12a-mediated miRNA knockout system to accurately generate loss-of-function microRNA (miRNA) mutants in ten different miRNA genes in rice. The findings of the study accelerate the understanding of miRNA functions, enhancing their application in genetic engineering research and efforts.

MiRNAs are small, non-coding RNAs in eukaryotes that regulate gene expression and play vital roles in plant development, stress responses, and metabolism. Traditional methods for studying miRNAs have been used to investigate miRNA function, but generating knockout mutants has been challenging due to the small size of miRNA genes. To achieve efficient genetic knockouts of miRNA genes, selecting the right CRISPR system is needed.

In this study, the researchers found that the Cas12a-generated T1 homozygous mutant seeds did not grow into any seedlings, while a few Cas9-generated T1 homozygous seeds developed elongated lateral roots without any shoot. Knockout mutants of nine miRNA genes in rice were also generated with the use of CRISPR-Cas12a technology. This data suggests that Cas12a is a more efficient tool for generating null alleles of miRNAs in plants. The researchers envision the use of CRISPR-Cas12a in creating genome-wide miRNA mutant libraries in plants in the future.

For more information, read the study from the Plant Biotechnology Journal.