The Need

CRISPR technology is an exciting tool for editing the genomes of live organisms. With CRISPR, researchers have replaced target genes with chosen alternative genes in species ranging from bacteria to live mice. The CRISPR system uses either the Cas9 or Cpf1 protein to bind a given DNA sequence and cleave it. The CRISPR/Cas9 system is the popular and widely used gold standard, but it suffers from some limitations relative to CRISPR/Cpf1. Cas9 requires two RNA molecules to activate DNA cleavage, whereas Cpf1 requires only one guide RNA (crRNA). Cleavage of DNA with Cas9 results in blunt ends, whereas Cpf1 produces sticky ends, the latter of which permits more specific genomic targeting. As a laboratory tool and as a potential therapeutic, there is a need to optimize the CRISPR system for efficiency and accuracy.

The Technology

Ohio State University researchers, led by Dr. Yizhou Dong, have created modified forms of the guide RNA (gRNA) that can improve Cpf1 nuclease activity, reduce toxicity, decrease off-target effects, and increase gRNA stability in the CRISPR/Cpf1 genome editing system. Because the CRISPR/Cpf1 system is simpler to the Cas9-based system, the Cpf1 system is expected to quickly gain popularity for both academic research and drug development. The modified forms of gRNA will facilitate the process and further improve the genome editing accuracy and efficiency of the system.

Commercial Applications

• Genome editing
• Genetic engineering
• Guide RNA

Benefits/ Advantages

• High gene editing efficiency
• Low off-target effects