Use of Cre/loxP recombination to swap cell binding motifs on the adenoviral capsid protein IX

Journal Article · Fri Nov 25 00:00:00 EST 2011 · Virology

DOI:https://doi.org/10.1016/j.virol.2011.09.004· OSTI ID:21587896

Poulin, Kathy L ^[1]; Tong, Grace; Vorobyova, Olga ^[1]; Pool, Madeline ^[1]; Kothary, Rashmi ^[1]

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1. Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, Ontario (Canada)

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. - Highlights: > We describe a method to grow virus lacking native tropism but containing novel cell-binding ligands. > Cre/loxP recombination was used to modify the adenovirus genome. > A targeting ligand present on capsid protein IX was removed or replaced using recombination. > Cre-loxP was also used to 'swap' the identity of the targeting ligand present on pIX.

• OSTI ID: 21587896
• Journal Information: Virology, Vol. 420, Issue 2; Other Information: DOI: 10.1016/j.virol.2011.09.004; PII: S0042-6822(11)00407-7; Copyright (c) 2011 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); ISSN 0042-6822
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
ADENOVIRUS
ANTIBODIES
FIBERS
GENE THERAPY
LIGANDS
LYSINE
NEOPLASMS
RECEPTORS
RECOMBINATION
TYROSINE
AMINO ACIDS
CARBOXYLIC ACIDS
DISEASES
HYDROXY ACIDS
MEDICINE
MEMBRANE PROTEINS
MICROORGANISMS
ONCOGENIC VIRUSES
ORGANIC ACIDS
ORGANIC COMPOUNDS
PARASITES
PROTEINS
THERAPY
VIRUSES