The amazing complexity of cellular networks is still not fully understood and therefore modifying a single gene to treat a patient can have various effects that need to be determined. Answering the questions about what is happening at the cellular level requires an untargeted and very sensitive approach. State-of-the-art, highly sensitive Zeno SWATH data-independent acquisition (DIA) can determine off- and on-target effects with unprecedented depth and short injection-to-injection times that allow for necessary throughput. Here, Hugo Gagnon will discuss experimental design, depth of analysis, target coverage and statistical challenges to help biopharmaceutical drug developers ensure safe and efficient gene therapy products.