Researchers from King Fahd University of Petroleum and Minerals and Texas Tech University reviewed the latest approaches used to minimize off-target effects in genome editing such as biased or unbiased off-target detection, cytosine or adenine base editors, prime editing, dCas9, Cas9 paired nickase, ribonucleoprotein delivery and truncated gRNAs. The review article is published in Cells.

According to the article, CRISPR-Cas9 has been the most popular genome editing tool because of its accuracy, efficiency, cost-effectiveness, and ease of use compared to others. However, CRISPR-Cas9 may cause off-target effects at the genome level and off-target detection tools are also necessary. The most preferred off-target detecting biased and unbiased methods presented were Elevation and GUIDE–seq, respectively. Furthermore, choosing Cas variants is also critical to reduce the off-target effects that depend on the nature of the experiment.


Read more in MDPI.



Figire 1: Effects of off-target mutation on animal and plant phenotype. Off-target causes genetic mutations. In clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system gRNA sometimes binds other than target loci, off-target site. It may activate the oncogenes that initiate tumor cell formation in the animal body or may change gene function that leads to undesirable phenotypic expression (sensitive to diseases) in plants. The figure has been created with