Improvement of lentiviral vector-mediated gene transduction by genetic engineering of the structural protein Pr55 Gag.
The lentiviral vector is a promising tool for human gene therapy because of its ability to transduce genes into many cell types. However, one of the technical problems associated with the lentiviral vector is that lentiviral titers in current production systems are relatively low compared with the other viral vectors. In this study, we provide genetic evidence that the attachment of heterologous myristoylation (myr) signals on the amino-terminus of human immunodeficiency virus type 1 Pr55(Gag) (Gag) can increase the viral yield up to 10-fold, leading to the enhancement of gene transduction in many cell lines. The myr signal Gag constructs behaved similarly to the wild-type Gag in targeting to detergent-resistant membrane compartments, Vps4-dependence for viral budding, and virion morphology. However, the myr signal Gag constructs showed improved oligomerization efficiency as measured by bioluminescence resonance energy transfer in living cells, contributing to increased viral production and efficient activation of the viral protease responsible for virion maturation. The genetically modified Gag represents the next generation lentiviral vector, and should contribute to the success of many lentiviral vector applications.
Aoki
T
T
AIDS Research Center, National Institute of Infectious Diseases, Tokyo, Japan.
Shimizu
S
S
Urano
E
E
Futahashi
Y
Y
Hamatake
M
M
Tamamura
H
H
Terashima
K
K
Murakami
T
T
Yamamoto
N
N
Komano
J
J
D016827:CD8 Antigens / Q000235:genetics / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D018613:Microscopy, Confocal; D011498:Protein Precursors / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods*
D016827:CD8 Antigens; D011498:Protein Precursors; C059767:p55 gag precursor protein, Human immunodeficiency virus 1