Genetically modified adenoviral vector with the protein transduction domain of Tat improves gene transfer to CAR-deficient cells.
The transduction efficiency of Ad (adenovirus) depends, to some extent, on the expression level of CAR (coxsackievirus and Ad receptor) of a target cell. The low level of CAR on the cell surface is a potential barrier to efficient gene transfer. To overcome this problem, PTD.AdeGFP (where eGFP is enhanced green fluorescent protein) was constructed by modifying the HI loop of Ad5 (Ad type 5) fibre with the Tat (trans-activating) PTD (protein transduction domain) derived from HIV. The present study showed that PTD.AdeGFP significantly improved gene transfer to multiple cell types deficient in expression of CAR. The improvement in gene transfer was not the result of charge-directed binding between the virus and the cell surface. Although PTD.AdeGFP formed aggregates, it infected target cells in a manner different from AdeGFP aggregates precipitated by calcium phosphate. In addition, PTD.AdeGFP was able to transduce target cells in a dynamin-independent pathway. The results provide some new clues as to how PTD.AdeGFP infects target cells. This new vector would be valuable in gene-function analysis and for gene therapy in cancer.
Liu
Shihai
S
Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an 710062, People's Republic of China.
Mao
Qinwen
Q
Zhang
Weifeng
W
Zheng
Xiaojing
X
Bian
Ye
Y
Wang
Dongyang
D
Li
Huijin
H
Chai
Lihong
L
Zhao
Junli
J
Xia
Haibin
H
D000260:Adenoviruses, Human / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D005347:Fibroblasts / Q000378:metabolism; D015696:Gene Products, tat / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D007668:Kidney / Q000166:cytology; D017434:Protein Structure, Tertiary / Q000235:genetics; D011991:Receptors, Virus / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*; D014471:Umbilical Veins / Q000166:cytology / Q000378:metabolism
C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D015696:Gene Products, tat; D011991:Receptors, Virus