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Recombinant Adeno-associated Viral (rAAV) Vectors: Production, Targeting, and Applications

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Janina Frisch

Janina Frisch

Saarland University, Center of Human and Molecular Biology, Biophysics Department

Janina Frisch (PhD) is a part of the Center of Human and Molecular Biology at the Biophysics Department of the Saarland University, Germany. From 2008 to 2011, she studied Applied Life Sciences at the University of Applied Sciences in Zweibruecken, Germany, where a guest research project at the Chinese University of Hong Kong was included. Thereafter, she attained her master’s degree in Biomedicine from Johannes Gutenberg University in Mainz, Germany (2011-2013). Her doctoral studies were performed from 2013 to 2016 at the Center of Experimental Orthopaedics, Saarland University Medical Center, Germany, wherein intensive research was done with rAAV vectors, to genetically modify stem cells and bone marrow aspirates for cartilage repair. Her thesis was awarded, and she also published several articles verifying the use of rAAV vectors to overexpress growth factors and to promote chondrogenic differentiation of progenitor cells. In her first postdoctoral phase, she continued working on rAAV vectors at the Paul Ehrlich Institute in Langen, Germany, where her research was focused on immunotherapy for cancer diseases, which involved the development of targeted rAAV vectors encoding for immune checkpoint inhibitors (2016-2018). At present, she is contributing to a Collaborative Research Center funded by the German Research Foundation by working on cardiovascular complications in chronic kidney disease. The laboratory is mainly performing redox research using a genetically encoded sensor with subcellular resolution in a broad application field. Establishing rAAV vector production and application is one of her main goals, along with inducing new and promising tools, to further increase the scope of research for the laboratory.

Collection Overview

Gene therapy is a consistently growing field of research that was experimentally initiated more than 45 years ago for a broad range of diseases, while clinical translation is relatively recent with the first commercial use being approved in 2005 in China. In this aspect, recombinant adeno-associated viral (rAAV) vectors serve as promising tools to induce genetic modifications with low immunogenicity and toxicity, while ensuring the achievement of efficient and long-term transgene expression. They are obtained from the replication-defective adeno-associated virus with basically 12 evident wildtype serotypes that are mainly entering the host cell via heparin sulfate proteoglycan (HSPG) and AAV receptors (AAVR). Recombinant AAV vectors can be either produced using a helper virus (such as adenovirus) or helper-free viruses, where the required adenoviral genes are provided via the helper-plasmid application. Several strategies for rAAV design are available. Naturally occurring single-stranded (ss) AAV vectors require self-priming induced second strand synthesis that leads to delayed transgene expression in the host cell. On the other hand, self-complementary (sc) vectors with dimeric inverted repeat genomes facilitate early transgene expression while the genomic packaging capacity is roughly half-sized. Furthermore, besides covering a broad tissue tropism by the wildtype serotypes, redirection/specific cellular targeting has gained much interest in recent years. For that, natural receptor binding is ablated, and peptide ligands are synthetically fused to the capsid proteins to allow target receptor binding and entry. Hence, methodologies and techniques described herein provide insights into the opportunities of rAAV vector production for different gene therapy approaches/aims.

Articles

Adeno-associated Virus-mediated Transgene Expression in Genetically Defined Neurons of the Spinal Cord
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Adeno-associated Virus-mediated Transgene Expression in Genetically Defined Neurons of the Spinal Cord

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Cited by 21

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2018

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