A high-capacity, capsid-modified hybrid adenovirus/adeno-associated virus vector for stable transduction of human hematopoietic cells.
To achieve stable gene transfer into human hematopoietic cells, we constructed a new vector, DeltaAd5/35.AAV. This vector has a chimeric capsid containing adenovirus type 35 fibers, which conferred efficient infection of human hematopoietic cells. The DeltaAd5/35.AAV vector genome is deleted for all viral genes, allowing for infection without virus-associated toxicity. To generate high-capacity DeltaAd5/35.AAV vectors, we employed a new technique based on recombination between two first-generation adenovirus vectors. The resultant vector genome contained an 11.6-kb expression cassette including the human gamma-globin gene and the HS2 and HS3 elements of the beta-globin locus control region. The expression cassette was flanked by adeno-associated virus (AAV) inverted terminal repeats (ITRs). Infection with DeltaAd5/35.AAV allowed for stable transgene expression in a hematopoietic cell line after integration into the host genome through the AAV ITR(s). This new vector exhibits advantages over existing integrating vectors, including an increased insert capacity and tropism for hematopoietic cells. It has the potential for stable ex vivo transduction of hematopoietic stem cells in order to treat sickle cell disease.
Shayakhmetov
Dmitry M
DM
Division of Medical Genetics, University of Washington, Seattle, Washington 98195, USA.
Carlson
Cheryl A
CA
Stecher
Hartmut
H
Li
Qiliang
Q
Stamatoyannopoulos
George
G
Lieber
André
A
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D004279:DNA, Viral; D000229:Dependovirus / Q000235:genetics* / Q000502:physiology; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics* / Q000502:physiology; D005914:Globins / Q000235:genetics; D049452:Green Fluorescent Proteins; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D020014:K562 Cells; D008164:Luminescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D011995:Recombination, Genetic; D020079:Terminal Repeat Sequences; D013997:Time Factors; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes; D016662:Virus Integration
D036022:Capsid Proteins; D004279:DNA, Viral; D008164:Luminescent Proteins; C018315:hexon capsid protein, Adenovirus; D049452:Green Fluorescent Proteins; D005914:Globins