De conclusie van genoemd artikel mbt off-target effecten van dna editing is in zoverre duidelijk dat zelfs base en/of prime editing leiden tot off target effecten die nader moeten worden onderzocht. De hamvraag is waar een FDA/EMA de streep zal trekken. Hoeveel off-target fouten gaat men accepteren voordat het definitieve groene licht wordt gegeven?
"The success of CRISPR/Cas9 to modify genome with unprecedented precision can be owed to its accuracy, efficiency, cost-effectiveness and ease of use than the classical gene-editing tools (ZFNs and TALEN). However, CRISPR/Cas tool causes deleterious off-target effects at the genomic level. Various techniques reported, including different bioinformatics approaches such as for in silico detection of off-target mutations along with improved on-target efficiency to ameliorate the off-target effects. Choosing an appropriate off-target detection tool, such as biased and unbiased with predictive on-target and off-target sites, is more critical than the CRISPR delivery system. Most preferred off-target detecting biased and unbiased methods are Elevation and GUIDE–seq, respectively. In gRNA modification, truncated sgRNA stipulates a simple way to reduce off-target effects and RNP based delivery is suitable in most cases to get higher on-target activity. Additionally, choosing Cas variants is also critical to reduce the off-target effects that depend on the nature of the experiment.
Additionally, choosing Cas variants is also critical to reduce off-target effects. Researchers are in the race of creating engineered Cas9 variants and new gene-targeting techniques with negligible off-target effects in mammalian cells for the improvement of therapeutic gene editing or genome surgery. The currently developed prime editing tool has future promises to treat genetic diseases with minimum off-target effects in human cells, but off-target effects also limit Prime editing applications. However, further study needs for the development of a super CRISPR system to treat genetic diseases in the healthcare system."