AAV - Projects

Title: Technology platform I: Creation of synthetic AAV vectors by DNA shuffling
Summary: We expanded our original AAV DNA family shuffling approach (Grimm et al., J. Virol. 2008) to a set of 12 natural AAV serotypes that we have available in the lab, and concurrently optimized and streamlined the entire protocol. As a result, we can now routinely create libraries of at least 1e6 diverse capsids in as little as 5 days. A video demonstration of the entire procedure was published in the Journal of Visualized Experiments and can be found here: Kienle & Senis et al., J. Vis. Exp. 2012.
People: Eike Kienle, Stefanie Grosse

Title: Technology platform II: Creation of synthetic AAV vectors by peptide display
Summary: Based on reports that insertion of 7-mer peptides into an exposed region of the AAV2 capsid, we genetically engineered 11 additional serotypes available in our lab to permit peptide display in these alternative capsids as well. Notably, we showed that this technology is fully compatible with our DNA shuffling platform (project #1). This results in unique AAV libraries with unmatched genetic diversities, several of which we currently screen on various target cells, including human PBMCs.
People: Eike Kienle, Kathleen Börner, Stefanie Grosse, Teresa Pankert

Title: Technology platform III: Creation of synthetic AAV vectors by rational assembly
Summary: Based on reports that insertion of 7-mer peptides into an exposed region of the AAV2 capsid, we genetically engineered 11 additional serotypes available in our lab to permit peptide display in these alternative capsids as well. Notably, we showed that this technology is fully compatible with our DNA shuffling platform (project #1). This results in unique AAV libraries with unmatched genetic diversities, several of which we currently screen on various target cells, including human PBMCs.
People: Rudolf Pisa, Gabriella Cotugno

Title: Comprehensive screening of novel AAV peptide display mutants
Summary: We inserted 9 re-targeting peptides into 12 distinct AAV serotypes, yielding a unique collection of 108 synthetic AAV capsids that we have screened on >30 cell types thus far, including various primary human cells. Our most impressive findings includes the isolation of a novel AAV capsid capable of mediating up to 100% transduction of various human T-cell lines which we currently engineer together with the group of Prof. Kräusslich as vector for anti-HIV shRNA delivery.
People: Marina Bechtle, Eike Kienle, Kathleen Börner, Elena Senis

Title: An AAV vector toolbox for iPSC reprogramming, marking and purging
Summary: We engineered a collection of double-stranded AAV vectors to express factors needed for reprogramming of somatic cells into induced pluripotent stem cells (iPSC). Moreover, we created gfp-expressing AAVs that can segregate fully reprogrammed iPSC from their ancestors based on differential miRNA expression. Finally, we also generated toxic AAVs for purging of naive iPSC from completely differentiated somatic cells as a safety measure for cell transplantation strategies.
People: Elena Senis, in collaboration with J. Utikal (Mannheim University Hospital) & Axel Schambach (Hannover Medical School)

Title: Targeted AAV vectors therapeutic or diagnostic RNAi approaches
Summary: We engineered a collection of double-stranded AAV vectors to express factors needed for reprogramming of somatic cells into induced pluripotent stem cells (iPSC). Moreover, we created gfp-expressing AAVs that can segregate fully reprogrammed iPSC from their ancestors based on differential miRNA expression. Finally, we also generated toxic AAVs for purging of naive iPSC from completely differentiated somatic cells as a safety measure for cell transplantation strategies.
People: Stefan Mockenhaupt, Nina Schürmann, Loredana Lazaru

Contact: E-Mail (Last update: 26/02/2012)