Project description:Recombinant viral vectors such as adeno-associated viruses (AAV) have emerged as a key tool in gene therapy applications. Despite extensive research efforts aimed at increasing production yields, the intricate molecular mechanisms that govern the synthesis of viral vectors within host cells remain largely unknown. The interplay between viral replication and cellular biosynthetic processes is complex and not fully understood, which poses challenges for effective manipulation. This study uses transcriptomics and RNA sequencing to investigate the cellular processes underlying viral vector production. Transcriptomics examines RNA molecules and their role in gene expression and has become invaluable in modern biological research. The advent of cost-effective, high-throughput sequencing technologies enables the comprehensive analysis of transcriptomes, providing valuable insights into gene regulation and cell signalling pathways. This study takes a strategic approach to deciphering the cellular mechanisms involved in viral vector production by utilising RNA-Seq data from a commercially available HEK293 suspension cell line. This study emphasises the importance of data-driven methodologies in advancing cell line development and optimising viral vector production.
Project description:Recombinant viral vectors such as adeno-associated viruses (AAV) have emerged as a key tool in gene therapy applications. Despite extensive research efforts aimed at increasing production yields, the intricate molecular mechanisms that govern the synthesis of viral vectors within host cells remain largely unknown. The interplay between viral replication and cellular biosynthetic processes is complex and not fully understood, which poses challenges for effective manipulation. This study uses transcriptomics and RNA sequencing to investigate the cellular processes underlying viral vector production. Transcriptomics examines RNA molecules and their role in gene expression and has become invaluable in modern biological research. The advent of cost-effective, high-throughput sequencing technologies enables the comprehensive analysis of transcriptomes, providing valuable insights into gene regulation and cell signalling pathways. This study takes a strategic approach to deciphering the cellular mechanisms involved in viral vector production by utilising RNA-Seq data from a commercially available HEK293 suspension cell line. This study emphasises the importance of data-driven methodologies in advancing cell line development and optimising viral vector production.
Project description:Recombinant adeno-associated viruses (rAAVs) are a cornerstone of modern gene therapy, offering precise delivery and stable transgene expression with minimal immunogenicity. Despite clinical advancements, large-scale rAAV production faces significant challenges such as low yield, high costs, and variability, hindering its scalability. Recent research suggests that miRNAs and snoRNAs can modulate viral replication, transcriptional regulation, and host defense mechanisms, making them attractive targets for improving rAAV production. This study examines the expression profiles of microRNAs (miRNAs) and small nucleolar RNAs (snoRNAs) during rAAV plasmid transfection and production in HEK293F cells. We used microarrays to detail differential expression during rAAV production versus mock transfection and basal expression and identified significantly up- and down-regulated small non-coding RNAs (ncRNAs).