One of the main bottlenecks to all gene therapy applications is the reliable and rapid synthesis of complex DNA constructs. This is especially true for exceedingly large constructs (> 5 kB), and ones with long-terminal repeats or complex genomic elements. After production of the base DNA construct, the delivery of large cargoes safely and effectively (> 5 kB) has proven challenging to the field, especially the clinical standard delivery vehicles of AAVs and LNPs. Ansa Bio and EndoFold sought to address these concerns in a proof-of-concept collaboration. The preliminary work features the rapid synthesis of a complex, full-length human dystrophin plasmid for use as a RNA cargo template for EndoFold’s proprietary gene delivery vehicles.
Key Takeaways
Rapid and reliable synthesis of large human dystrophin constructs (UTRs and ~23 kB) within ~20 business days from sequence submission, whereas other DNA suppliers could not support the same timely delivery.
Cost effective and retained complexity in the full-length dystrophin plasmid permits immediate use of full-length dystrophin as a cargo with EndoFold’s proprietary, AI+physics designed delivery vehicles.
Robust expression from the dystrophin construct when placed in human cells.