Pharmacy researchers engineer RNA system to maximize genome editing

05/15/2017

CRISPR (clustered regularly interspaced short palindromic repeats) allows researchers to precisely edit targeted DNA sequences and genomes. Yizhou Dong, PhD, assistant professor at The Ohio State University College of Pharmacy Division of Pharmaceutics and Pharmaceutical Chemistry has engineered an RNA system that has substantially improved gene cutting and was recently published in Nature Biomedical Engineering.

Cpf1, reported previously, is a type V CRISPR-Cas effector endonuclease. The Cpf1 system exhibits gene editing activity in human cells through a single RNA-guided approach. Although this system offers a new tool, low efficiency is one of the key challenges for its clinical uses. To address this issue, Dong and co-workers applied an engineering approach to the Cpf1 system.

“We identified the optimal Cpf1 guide RNA and messenger RNA, and then combined them to improve the gene-cutting efficiency in human cells,” said Dong. “I hope these results help us further understand the Cpf1 system and provide useful design criteria for maximizing genome editing efficiency in biological and therapeutic applications.” 

The engineered system was successfully tested on different members of the Cpf1 family. Dong’s findings show that the modified system did improve efficiency in three cell lines and for three target genomic sites.

The results of Dong’s research demonstrate the broad applications for the new RNA system. For example, if there is a mutation in a genome, engineered Cpf1 system facilitate researchers to cut the mutated site so that the gene can be edited more effectively.

Other authors in the article include Bin Li, Weiyu Zhao, Xiao Luo, Xinfu Zhang, Chenglong Li, and Chunxi Zeng. Prof. Feng Zhang and Mr. Bernd Zetsche at the Broad Institute generously provided reagents and technical assistance.