Location: Nantes, France
Leader for 20 years on viral vector production and translational research for Gene Therapy, the UMR1089 Inserm Research department (Gene Therapy Laboratory) at Nantes (France) is looking for a M2 student.
Impact of new AAV genome extremities on the cell sensing and on the efficiency of gene transfer
Adeno-Associated Viral Virus (AAV) are one of the most suitable viral vectors for gene delivery in vivo. Encouraging preclinical data using AAV vectors have been followed by several successful gene therapy clinical trials and by the marketing authorization of two AAV-based products, Glybera® and Luxturna®. The scientific community is now facing news issues addressing systemic diseases such as Duchenne muscular dystrophy. In addition to manufacturing limitations, injecting more than 1014 AAV particles per kg may trigger unwanted side effects such as an immune response against the viral capsids. AAV particle and genome can also be sensed by the targeted cells and lead to a decrease in the expression of the gene of interest. Thus, it would appear essential to optimize both AAV capsid and recombinant genome in order to reduce the therapeutic doses and improve the efficiency and safety of these vectors.The AAV genome is a single-stranded DNA of 4.7 kb composed of three open reading frames, rep, cap and AAP. To design a recombinant AAV vector, these ORFs are replaced by the therapeutic gene. The only viral sequences that remain in the vector genome are the ITR (inverted terminal repeats). ITR are 145 base-sequences comprising palindromic regions that can form a T-shaped structure (Figure). These sequences can be recognized by the cellular sensing pathways after transduction.
In our team, we have designed new ITR sequences in order to avoid this cellular response. The M2 student will generate AAV vectors from these new sequences, test these vectors on primary cells and implement assays so as to determine which restriction pathway (Toll-like receptor, DNA damage response) is bypassed by the modified terminal repeats. The candidate must be interested in molecular biology, virology and cellular sensing pathways. He will take part of a more general project on the optimization of recombinant AAV genomes. Qualities such a rigor, organization and motivation will be appreciated.
Contact : firstname.lastname@example.org
The UMR1089 Inserm Research department (Gene Therapy Laboratory) is member of West Biotherapy.
#gene therapy, #adeno-associated virus, #viral vectors, #inverted terminal repeats, #cell sensing, #gene regulation