title: In vitro incorporation of a cell-binding protein to a lentiviral vector using an engineered split intein enables targeted delivery of genetic cargo.
abstract: Gene therapy represents a promising therapeutic paradigm for addressing many disorders, but the absence of a vector that can be robustly and reproducibly functionalized with cell-homing functionality to mediate the delivery of genetic cargo specifically to target cells following systemic administration has stood as a major impediment. In this study, a high-affinity protein-protein pair comprising a splicing-deficient naturally split intein was used as molecular Velcro to append a HER2/neu-binding protein (DARPin) onto the surface of a binding-deficient, fusion-competent lentivirus. HER2/neu-specific lentiviruses created using this in vitro pseudotyping approach were able to deliver their genetic reporter cargo specifically to cells that express the target receptor at high levels in a co-culture. We envision that the described technology could provide a powerful, broadly applicable platform for the incorporation of cell-targeting functionality onto viral vectors.
authors.affiliation: Department of Microbial and Molecular Pathogenesis, Texas A&M Health Science Center, College Station, Texas, 77843.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Chamoun-Emanuelli
authors.affiliation: Department of Veterinary Physiology and Pharmacology, Texas A&M University, College Station, Texas.
authors.forename: Gus
authors.initials: G
authors.lastname: Wright
authors.affiliation: Department of Veterinary Physiology and Pharmacology, Texas A&M University, College Station, Texas.
authors.forename: Smith
authors.initials: S
authors.lastname: Roger
authors.affiliation: Division of Medical Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Münch
authors.affiliation: Division of Medical Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: Christian J
authors.initials: CJ
authors.lastname: Buchholz
authors.affiliation: Department of Microbial and Molecular Pathogenesis, Texas A&M Health Science Center, College Station, Texas, 77843. zchen4@tamu.edu.
authors.forename: Zhilei
authors.initials: Z
authors.lastname: Chen
mesh_terms: D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism*; D016503:Drug Delivery Systems; D015316:Genetic Therapy / Q000379:methods*; D016086:Lentivirus / Q000235:genetics* / Q000502:physiology*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism; D011991:Receptors, Virus / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D053585:Virus Attachment
chemical_list: D002352:Carrier Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D011991:Receptors, Virus; D011994:Recombinant Proteins; C508053:ERBB2 protein, human
keywords: HER2/neu; gene therapy; lentivirus