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This research aims to evaluate the efficacy of CROFT-Seq in identifying off-target sites of CRISPR-Cas9 nucleases.

April 25, 2026Synthetic BiologyOpen Access

Off-target detection of CRISPR-Cas9 nuclease in vitro with CROFT-Seq

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Randomized trial reveals a rapid method for detecting off-target sites in genome editing, suggesting improved safety for the technology.

This research aims to evaluate the efficacy of CROFT-Seq in identifying off-target sites of CRISPR-Cas9 nucleases.
Developed CROFT-Seq for genome-wide detection of Cas9 off-target sites in vitro.
Compared performance with existing in vitro detection methods.
Validated top-ranked off-target sites in cellular models.
CROFT-Seq effectively identified off-target sites comparable to standard methods.
A high percentage of top-ranked off-target sites confirmed in cells, demonstrating strong predictive accuracy.

Toliusis et al. (2026) studied this question.

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