DisTAL-Seq: A NEW METHOD TO PROFILE TALEN OFF-TARGETS — PUBLISHED IN MOLECULAR THERAPY: NUCLEIC ACIDS

Programmable nucleases have transformed genome editing, enabling precise DNA modification for both research and therapeutic applications. However, ensuring that these tools cut only at their intended target—and not elsewhere in the genome—remains a key challenge, particularly for clinically relevant platforms where accurate off-target detection is essential.

In our latest collaboration with the Genome Engineering and Measurement Lab (GEML) and Allogene Therapeutics, we introduce DisTAL-Seq, a method that enables genome-wide detection of TALEN-induced DNA double strand breaks directly in human cells. The experimental work was led by research technician Lena Kobel. This approach builds on the principles of DISCOVER-Seq and incorporates analysis logic tailored to TALEN binding architecture, including variable repeat specificity, cleavage offset, and dimerization behavior.

Using DisTAL-Seq, we identified and validated editing sites across different TALEN designs and primary human T-cell donors, providing a systematic view of TALEN specificity in clinically relevant contexts.

These results expand the genome-wide profiling approaches developed at GEML and provide a framework for evaluating the safety and performance of genome editing nucleases as they move towards therapeutic applications.

For more detail, check out our paper in Molecular Therapy: Nucleic Acids.