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

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

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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DOI

https://doi.org/10.3390/ijms20225702

URN

urn:nbn:de:0070-pub-29391289

Autor*in

Feiner, Rebecca^UniBi; Teschner, Julian^UniBi; Teschner, Kathrin^UniBi; Radukic, Marco^UniBi; Baumann, Tobias; Hagen, Sven; Hannappel, Yvonne^UniBi; Biere, Niklas^UniBi; Anselmetti, Dario^UniBi ; Arndt, Katja M.; Müller, Kristian^UniBi

Einrichtung

Technische Fakultät > AG Zelluläre und molekulare Biotechnologie

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

Ausgabe

Art.-Nr.

5702

Urheberrecht / Lizenzen

Creative Commons Namensnennung 4.0 International Public License (CC-BY 4.0)

ISSN

1422-0067

eISSN

1422-0067

Finanzierungs-Informationen

Open-Access-Publikationskosten wurden durch die Deutsche Forschungsgemeinschaft und die Universität Bielefeld gefördert.

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. https://doi.org/10.3390/ijms20225702

Feiner, Rebecca, Teschner, Julian, Teschner, Kathrin, Radukic, Marco, Baumann, Tobias, Hagen, Sven, Hannappel, Yvonne, 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.

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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