Tonight, I watched the London Calling 2024 session by Kathleen Zeglinski from the Walter and Eliza Hall Institute of Medical Research in Australia. The session title was “Quality Control of Gene Therapy Vectors using Nanopore Direct RNA Sequencing.” Zeglinski explained that lentiviruses are a type of RNA viruses that can integrate into the genome. Lentiviruses can be used for gene therapy and require lentiviral vector quality control. Long-read sequencing could be used to identify truncated RNA or isoforms. Direct RNA sequencing was used because it provides an unbiased view. Plots of coverage changes can be used to identify cryptic elements. The team artificially polyadenylated sequences to reveal hairpin-associated truncations. The results of these experiments were used to quantify the amount of complete RNA and what to address when optimizing the vectors. The team also used the Induro Reverse Transcriptase, which improved the sequencing coverage. Zeglinski shared examples of improving vectors and some of the effects of changes, concluding that vector improvement is challenging! They noted that sequencing each vector design helps identify potential problems that were created. I had not considered direct RNA sequencing for optimizing lentiviral gene therapy vectors!
