rAAV Engineering for Capsid-Protein Enzyme Insertions and Mosaicism Reveals Resilience to Mutational, Structural and Thermal Perturbations

Feiner R, Teschner J, Teschner K, Radukic M, Baumann T, Hagen S, Hannappel Y, Biere N, Anselmetti D, Arndt KM, Müller K (2019)
International Journal of Molecular Sciences 20(22): 5702.

Zeitschriftenaufsatz | Veröffentlicht| Englisch
 
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Abstract / Bemerkung
Recombinant adeno-associated viruses (rAAV) provide outstanding options for customization and superior capabilities for gene therapy. To access their full potential, facile genetic manipulation is pivotal, including capsid loop modifications. Therefore, we assessed capsid tolerance to modifications of the structural VP proteins in terms of stability and plasticity. Flexible glycine-serine linkers of increasing sizes were, at the genetic level, introduced into the 587 loop region of the VP proteins of serotype 2, the best studied AAV representative. Analyses of biological function and thermal stability with respect to genome release of viral particles revealed structural plasticity. In addition, insertion of the 29 kDa enzyme β-lactamase into the loop region was tested with a complete or a mosaic modification setting. For the mosaic approach, investigation of VP2 trans expression revealed that a Kozak sequence was required to prevent leaky scanning. Surprisingly, even the full capsid modification with β-lactamase allowed for the assembly of capsids with a concomitant increase in size. Enzyme activity assays revealed lactamase functionality for both rAAV variants, which demonstrates the structural robustness of this platform technology.
Stichworte
adeno-associated-virus; β-lactamase; inverted terminal repeat (ITR); loop modification; capsid stability
Erscheinungsjahr
2019
Zeitschriftentitel
International Journal of Molecular Sciences
Band
20
Ausgabe
22
Art.-Nr.
5702
eISSN
1422-0067
Finanzierungs-Informationen
Article Processing Charge funded by the Deutsche Forschungsgemeinschaft and the Open Access Publication Fund of Bielefeld University.
Page URI
https://pub.uni-bielefeld.de/record/2939128

Zitieren

Feiner R, Teschner J, Teschner K, et al. rAAV Engineering for Capsid-Protein Enzyme Insertions and Mosaicism Reveals Resilience to Mutational, Structural and Thermal Perturbations. International Journal of Molecular Sciences. 2019;20(22): 5702.
Feiner, R., Teschner, J., Teschner, K., Radukic, M., Baumann, T., Hagen, S., Hannappel, Y., et al. (2019). rAAV Engineering for Capsid-Protein Enzyme Insertions and Mosaicism Reveals Resilience to Mutational, Structural and Thermal Perturbations. International Journal of Molecular Sciences, 20(22), 5702. doi:10.3390/ijms20225702
Feiner, R., Teschner, J., Teschner, K., Radukic, M., Baumann, T., Hagen, S., Hannappel, Y., Biere, N., Anselmetti, D., Arndt, K. M., et al. (2019). rAAV Engineering for Capsid-Protein Enzyme Insertions and Mosaicism Reveals Resilience to Mutational, Structural and Thermal Perturbations. International Journal of Molecular Sciences 20:5702.
Feiner, R., et al., 2019. rAAV Engineering for Capsid-Protein Enzyme Insertions and Mosaicism Reveals Resilience to Mutational, Structural and Thermal Perturbations. International Journal of Molecular Sciences, 20(22): 5702.
R. Feiner, et al., “rAAV Engineering for Capsid-Protein Enzyme Insertions and Mosaicism Reveals Resilience to Mutational, Structural and Thermal Perturbations”, International Journal of Molecular Sciences, vol. 20, 2019, : 5702.
Feiner, R., Teschner, J., Teschner, K., Radukic, M., Baumann, T., Hagen, S., Hannappel, Y., Biere, N., Anselmetti, D., Arndt, K.M., Müller, K.: rAAV Engineering for Capsid-Protein Enzyme Insertions and Mosaicism Reveals Resilience to Mutational, Structural and Thermal Perturbations. International Journal of Molecular Sciences. 20, : 5702 (2019).
Feiner, Rebecca, Teschner, Julian, Teschner, Kathrin, Radukic, Marco, Baumann, Tobias, Hagen, Sven, Hannappel, Yvonne, Biere, Niklas, Anselmetti, Dario, Arndt, Katja M., and Müller, Kristian. “rAAV Engineering for Capsid-Protein Enzyme Insertions and Mosaicism Reveals Resilience to Mutational, Structural and Thermal Perturbations”. International Journal of Molecular Sciences 20.22 (2019): 5702.
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2019-11-22T10:14:51Z
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