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Combined Genetic and Chemical Capsid Modifications of Adenovirus-Based Gene Transfer Vectors for Shielding and Targeting.
J Vis Exp. 2018 10 26JV

Abstract

Adenovirus vectors are potent tools for genetic vaccination and oncolytic virotherapy. However, they are prone to multiple undesired vector-host interactions, especially after in vivo delivery. It is a consensus that the limitations imposed by undesired vector-host interactions can only be overcome if defined modifications of the vector surface are performed. These modifications include shielding of the particles from unwanted interactions and targeting by the introduction of new ligands. The goal of the protocol presented here is to enable the reader to generate shielded and, if desired, retargeted human adenovirus gene transfer vectors or oncolytic viruses. The protocol will enable researchers to modify the surface of adenovirus vector capsids by specific chemical attachment of synthetic polymers, carbohydrates, lipids, or other biological or chemical moieties. It describes the cutting-edge technology of combined genetic and chemical capsid modifications, which have been shown to facilitate the understanding and overcoming of barriers for in vivo delivery of adenovirus vectors. A detailed and commented description of the crucial steps for performing specific chemical reactions with biologically active viruses or virus-derived vectors is provided. The technology described in the protocol is based on the genetic introduction of (naturally absent) cysteine residues into solvent-exposed loops of adenovirus-derived vectors. These cysteine residues provide a specific chemical reactivity that can, after production of the vectors to high titers, be exploited for highly specific and efficient covalent chemical coupling of molecules from a wide variety of substance classes to the vector particles. Importantly, this protocol can easily be adapted to perform a broad variety of different (non-thiol-based) chemical modifications of adenovirus vector capsids. Finally, it is likely that non-enveloped virus-based gene transfer vectors other than adenovirus can be modified from the basis of this protocol.

Authors+Show Affiliations

Center of Biomedical Education and Research, University Witten/Herdecke.Center of Biomedical Education and Research, University Witten/Herdecke.Center of Biomedical Education and Research, University Witten/Herdecke; florian.kreppel@uni-wh.de.

Pub Type(s)

Journal Article
Video-Audio Media

Language

eng

PubMed ID

30417881

Citation

Jönsson, Franziska, et al. "Combined Genetic and Chemical Capsid Modifications of Adenovirus-Based Gene Transfer Vectors for Shielding and Targeting." Journal of Visualized Experiments : JoVE, 2018.
Jönsson F, Hagedorn C, Kreppel F. Combined Genetic and Chemical Capsid Modifications of Adenovirus-Based Gene Transfer Vectors for Shielding and Targeting. J Vis Exp. 2018.
Jönsson, F., Hagedorn, C., & Kreppel, F. (2018). Combined Genetic and Chemical Capsid Modifications of Adenovirus-Based Gene Transfer Vectors for Shielding and Targeting. Journal of Visualized Experiments : JoVE, (140). https://doi.org/10.3791/58480
Jönsson F, Hagedorn C, Kreppel F. Combined Genetic and Chemical Capsid Modifications of Adenovirus-Based Gene Transfer Vectors for Shielding and Targeting. J Vis Exp. 2018 10 26;(140) PubMed PMID: 30417881.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Combined Genetic and Chemical Capsid Modifications of Adenovirus-Based Gene Transfer Vectors for Shielding and Targeting. AU - Jönsson,Franziska, AU - Hagedorn,Claudia, AU - Kreppel,Florian, Y1 - 2018/10/26/ PY - 2018/11/13/entrez PY - 2018/11/13/pubmed PY - 2019/4/2/medline JF - Journal of visualized experiments : JoVE JO - J Vis Exp IS - 140 N2 - Adenovirus vectors are potent tools for genetic vaccination and oncolytic virotherapy. However, they are prone to multiple undesired vector-host interactions, especially after in vivo delivery. It is a consensus that the limitations imposed by undesired vector-host interactions can only be overcome if defined modifications of the vector surface are performed. These modifications include shielding of the particles from unwanted interactions and targeting by the introduction of new ligands. The goal of the protocol presented here is to enable the reader to generate shielded and, if desired, retargeted human adenovirus gene transfer vectors or oncolytic viruses. The protocol will enable researchers to modify the surface of adenovirus vector capsids by specific chemical attachment of synthetic polymers, carbohydrates, lipids, or other biological or chemical moieties. It describes the cutting-edge technology of combined genetic and chemical capsid modifications, which have been shown to facilitate the understanding and overcoming of barriers for in vivo delivery of adenovirus vectors. A detailed and commented description of the crucial steps for performing specific chemical reactions with biologically active viruses or virus-derived vectors is provided. The technology described in the protocol is based on the genetic introduction of (naturally absent) cysteine residues into solvent-exposed loops of adenovirus-derived vectors. These cysteine residues provide a specific chemical reactivity that can, after production of the vectors to high titers, be exploited for highly specific and efficient covalent chemical coupling of molecules from a wide variety of substance classes to the vector particles. Importantly, this protocol can easily be adapted to perform a broad variety of different (non-thiol-based) chemical modifications of adenovirus vector capsids. Finally, it is likely that non-enveloped virus-based gene transfer vectors other than adenovirus can be modified from the basis of this protocol. SN - 1940-087X UR - https://www.unboundmedicine.com/medline/citation/30417881/Combined_Genetic_and_Chemical_Capsid_Modifications_of_Adenovirus_Based_Gene_Transfer_Vectors_for_Shielding_and_Targeting_ L2 - https://doi.org/10.3791/58480 DB - PRIME DP - Unbound Medicine ER -