Nucleic acids researchNov 11, 2024

CRISPRoffT: Complete database of unintended CRISPR/Cas gene editing targets

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗OA ↗

Updated Mar 28, 2026

Abstract

CRISPRoffT features data from 74 studies with 8840 validated off-targets.

A comprehensive repository of off-target regions has been developed to support gene-editing therapies.
CRISPRoffT integrates off-target analysis from various technologies, encompassing 29 prediction techniques.
The resource includes data on 368 guide sequences and 226,164 potential guide and off-target pairs.
It features off-target data from 85 different Cas/guide RNA combinations tested in 34 human and mouse cell lines.
Comparative analyses for individual guide sequences, genes, and techniques are available to inform experimental design.

Simplified

The CRISPR (clustered regularly interspaced short palindromic repeats)/Cas (CRISPR-associated protein) programmable nuclease system continues to evolve, with in vivo therapeutic gene editing increasingly applied in clinical settings. However, off-target effects remain a significant challenge, hindering its broader clinical application. To enhance the development of gene-editing therapies and the accuracy of prediction algorithms, we developed CRISPRoffT (https://ccsm.uth.edu/CRISPRoffT/). Users can access a comprehensive repository of off-target regions predicted and validated by a diverse range of technologies across various cell lines, Cas enzyme variants, engineered sgRNAs (single guide RNAs) and CRISPR editing systems. CRISPRoffT integrates results of off-target analysis from 74 studies, encompassing 29 experimental prediction techniques, 368 guide sequences, 226 164 potential guide and off-target pairs and 8840 validated off-targets. CRISPRoffT features off-target data from different CRISPR approaches (knockout, base editing and prime editing) applied under diverse experimental conditions, including 85 different Cas/guide RNA (gRNA) combinations used across 34 different human and mouse cell lines. CRISPRoffT provides results of comparative analyses for individual guide sequences, genes, cell types, techniques and Cas/gRNA combinations under different conditions. CRISPRoffT is a unique resource providing valuable insights that facilitate the safety-driven design of CRISPR-based therapeutics, inform experimental design, advance the development of computational off-target prediction algorithms and guide RNA design algorithms.

Key numbers

226 164

Total Off-Target Pairs

Total number of potential guide and off-target pairs in CRISPRoffT.

8840

Validated Off-Targets

Total number of experimentally validated off-targets included in the database.

Experimental Prediction Technologies

Number of different experimental prediction technologies used in the database.

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CRISPRoffT: Complete database of unintended CRISPR/Cas gene editing targets

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You found one interesting study. We’ll send the next 7.

what lands in your inbox each week:

Recent issues from the crispr gene editing brief