Feiner, Rebecca; Teschner, Julian; Teschner, Kathrin; Radukic, Marco; Baumann, Tobias; Hagen, Sven; Hannappel, Yvonne; Biere, Niklas; Anselmetti, Dario ; Arndt, Katja M.; Müller, Kristian: rAAV Engineering for Capsid-Protein Enzyme Insertions and Mosaicism Reveals Resilience to Mutational, Structural and Thermal Perturbations. In: International Journal of Molecular Sciences. Jg.20 H. 22. 2019
Inhalt
- Results
- Modifications of ITR and RepCap Plasmids Are Compatible with rAAV Production
- Systematic Variation of Loop Modifications Shows A Complex Pattern of Stability and Transduction Efficiency
- Mosaic rAAVs with a 29 kDa -Lactamase at Position 578 in VP2 Require a Kozak Consensus Sequence
- Fully Lactamase-Decorated rAAV Capsids Can Be Produced and Show Enzyme Activity
- Discussion
- Materials and Methods
- Construction of Plasmids
- Cell Culture
- Viral Particle Production
- Purification of Viral Particles
- SDS-PAGE and Western Blot Analysis
- Determination of Genomic Titers
- Transducing Titer Assays
- Transmission Electron Microscopy
- Atomic Force Microscopy
- AAV Stability Assay
- -Lactamase Activity Assays
- Statistical Analysis and Reproducibility
- References