Off-target genomic DNA sites in human B cells

Off-target genomic DNA sites in human B cells refer to unintended locations within the genome where gene-editing technologies, such as CRISPR-Cas9, induce double-strand breaks or chemical modifications. These sites typically possess sequence homology to the intended target site, leading to non-specific binding and activity of the editing machinery (Fu et al., 2013, Nature Biotechnology). In the context of B cells, which are frequently targeted for ex vivo therapies like CAR-T or CAR-B cell engineering, off-target effects pose significant risks. These risks include the potential for insertional mutagenesis or chromosomal translocations that could lead to oncogenic transformation (Kosicki et al., 2018, Nature Biotechnology). Monitoring and minimizing these off-target events is a critical component of safety assessments for genetic medicines. While not a therapeutic target themselves, these sites represent a major hurdle in the development of precise genomic interventions (Tsai et al., 2015, Nature Biotechnology). Advanced sequencing techniques like GUIDE-seq and CIRCLE-seq are employed to identify these sites and assess the precision of the editing platform. Minimizing off-target activity is essential for the clinical translation of gene-edited B-cell products to ensure patient safety and therapeutic efficacy.

Other names

Off-target effectsUnintended genomic modificationsNon-specific DNA cleavage sitesCRISPR off-targetsGenomic off-targets

Unintended DNA cleavage or modification at sequences with high homology to the target site