title: Genetically modified adenoviral vector with the protein transduction domain of Tat improves gene transfer to CAR-deficient cells.
abstract: 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.
authors: 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.
authors: Shihai
authors: S
authors: Liu
authors: Qinwen
authors: Q
authors: Mao
authors: Weifeng
authors: W
authors: Zhang
authors: Xiaojing
authors: X
authors: Zheng
authors: Ye
authors: Y
authors: Bian
authors: Dongyang
authors: D
authors: Wang
authors: Huijin
authors: H
authors: Li
authors: Lihong
authors: L
authors: Chai
authors: Junli
authors: J
authors: Zhao
authors: Haibin
authors: H
authors: Xia
mesh_terms: 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
chemical_list: C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D015696:Gene Products, tat; D011991:Receptors, Virus