Transfection of the urothelium was the most daunting challenge to intravesical gene therapy, independent of the presence of appropriate cellular receptors for the selected vector. The urothelium is protected by a glycocalyx, including the glycosaminoglycan (GAG) layer, which serves various functions, including the prevention of bladder infections and in the case of gene therapy, the prevention of bladder infection by viral vectors (). One common approach is to use chemicals to disrupt the urothelial barrier to enhance viral uptake. Initial success was reported using ethanol and acetone. However, adverse effects including hemorrhagic cystitis and stone formation limited its utility. In pursuit of a safer and more clinically useful transduction-enhancing agent the detergent Big CHAP [N,N0 -bis(3-D-gluconamidopropyl)cholamide] was evaluated in intravesical gene therapy (). Ironically, the commercially available Big CHAP’s yielded inconsistent results, and it was an impurity in one of the formularies that facilitated effective gene transfer. This impurity was the third identified, hence it was named Syn3. Syn3 is an anionic detergent which was identified as a contaminant in only certain lots of Big CAHP. Syn3 enhanced transgene expression in urothelium and resembled BigCHAP except for a substitution of a second cholyl group for one of the gluconic acid moieties. A more soluble form of Syn3 was synthesized at Canji by substituting disaccharide lactobionic acid for the gluconic acid to increase its solubility (). This discovery of Syn3 laid the foundation for the development of adenoviral interferon gene therapy for BLCA (, ).Interferon gene therapy for bladder cancer with nadofaragene firadenovec, Adstiladrin (nadofaragene firadenovec-vncg) is a gene therapy used to treat high-risk Bacillus CalmetteGuérin (BCG)-unresponsive non-muscle invasive bladder cancer. Includes Adstiladrin side effects, interactions and indications., Ferring’s novel adenovirus vector-based gene therapy Adstiladrin® (nadofaragene firadenovec-vncg) is the first gene therapy approved for bladder cancer Efficacy and safety of Adstiladrin supported by Phase 3 results demonstrating that more than half of patients (51% of CIS ± Ta/T1 cohort) achieved a complete response (CR) at three months and of these, 46% continued to remain free of high .