Use of Cre/loxP recombination to swap cell binding motifs on the adenoviral capsid protein IX
| Autor: | Olga Vorobyova, Rashmi Kothary, Kathy L. Poulin, Robin J. Parks, Grace Tong, Madeline Pool |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Coxsackie and Adenovirus Receptor-Like Membrane Protein
Recombinant Fusion Proteins Amino Acid Motifs Virus Attachment Cre recombinase Biology GPI-Linked Proteins Ligands Virus Replication Virus Receptor tyrosine kinase Adenoviridae Cell Line 03 medical and health sciences Gene therapy Antigens CD Virology Animals Humans Coding region Adenovirus Polylysine Virus retargeting Amino Acid Sequence Receptor trkA 030304 developmental biology Recombination Genetic 0303 health sciences Protein IX 030302 biochemistry & molecular biology Molecular biology Fusion protein Protein Structure Tertiary Rats 3. Good health HEK293 Cells Capsid Tissue tropism biology.protein Receptors Virus Capsid Proteins Infection Cell Adhesion Molecules Oncovirus Protein Binding |
| Zdroj: | Virology. 420(2):146-155 |
| ISSN: | 0042-6822 |
| DOI: | 10.1016/j.virol.2011.09.004 |
| Popis: | We used Cre/loxP recombination to swap targeting ligands present on the adenoviral capsid protein IX (pIX). A loxP-flanked sequence encoding poly-lysine (pK—binds heparan sulfate proteoglycans) was engineered onto the 3′-terminus of pIX, and the resulting fusion protein allowed for routine virus propagation. Growth of this virus on Cre-expressing cells removed the pK coding sequence, generating virus that could only infect through alternative ligands, such as a tyrosine kinase receptor A (TrkA)-binding motif engineered into the capsid fibre protein for enhanced infection of neuronal cells. We used a similar approach to swap the pK motif on pIX for a sequence encoding a single-domain antibody directed towards CD66c for targeted infection of cancer cells; Cre-mediated removal of the pK-coding sequence simultaneously placed the single-domain antibody coding sequence in frame with pIX. Thus, we have developed a simple method to propagate virus lacking native viral tropism but containing cell-specific binding ligands. |
| Databáze: | OpenAIRE |
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