Seed World

New Method Developed to Comprehensively Map the Off-Target Activity of CRISPR-Cas9 Across Entire Genomes


Caribou Biosciences, Inc., a leading company in CRISPR-Cas genome engineering, and DuPont Pioneer announced the Nature Methods publication describing SITE-Seq, a new method for comprehensively mapping CRISPR-Cas9 cleavage sites across the genome of any organism. When compared against other off-target analysis techniques, SITE-Seq detected potential off-target cleavage sites with a higher degree of resolution. This study has significant implications for the commercialization of products developed with CRISPR-Cas9 technology, as robust analytical tools are necessary to ensure the effective use of this gene-editing tool.

The full study, entitled “Mapping the genomic landscape of CRISPR,Cas9 cleavage,” was published May 1 in Nature Methods and details the biochemical method developed by Caribou and DuPont Pioneer scientists to comprehensively map CRISPR-Cas9 cleavage sites within the genome. Importantly, this information can be used to precisely measure the off-target effects of CRISPR-Cas9 genome editing in cell-based applications. Overall, this method will allow researchers to more effectively select CRISPR-Cas9 targets and delivery methods with maximum accuracy and specificity, thereby overcoming some of the concerns associated with potential off-target effects of CRISPR-Cas9.

This new method is expected to have broad applicability as it can be used to comprehensively detect CRISPR-Cas9 cleavage sites across the genome of any organism. The SITE-Seq method, in conjunction with cell-based validation methods, will constitute a robust pipeline for selecting CRISPR-Cas9 reagents for research or product development.

“This research marks a significant step towards the confirmation of CRISPR-Cas9 as a safe and effective gene-editing tool. By providing researchers with a way to accurately map the genomic landscape of CRISPR,Cas9 activity, we can reduce the occurrence of off-target effects by selecting for targets with the highest possible activity and specificity,” says Rachel Haurwitz, president and CEO of Caribou. “We are delighted at this latest demonstration of the valuable collaboration between Caribou and DuPont Pioneer scientists who are advancing the development of CRISPR-Cas9 applications.”

“The SITE-Seq research is an excellent proof point of productive collaboration between Caribou and DuPont Pioneer for the advancement of CRISPR-Cas technology,” says Neal Gutterson, vice president, Research & Development at DuPont Pioneer.