Construction and Gene Expression Analysis of a Single-Stranded DNA Minivector Based on an Inverted Terminal Repeat of Adeno-Associated Virus
Construction and Gene Expression Analysis of a Single-Stranded DNA Minivector Based on an Inverted Terminal Repeat of Adeno-Associated Virus
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基于腺相关病毒反向末端重复的单链DNA微型载体的构建和基因表达分析
DOI：
10.1007/s12033-014-9832-3
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发表时间：
2015
影响因子：
2.6
通讯作者：
Chun Zhang
中科院分区：
文献类型：
--
作者：
H. Ping;Xiaomei Liu;Dongqin Zhu;Tai;Chun Zhang
The plasmid vectors currently used for nonviral gene transfer have the disadvantage of carrying a bacterial backbone and an antibiotic resistance gene, which may cause side effects. The adeno-associated virus (AAV) genome is a linear single-stranded DNA (ssDNA) molecule with palindromic inverted terminal repeat (ITR) sequences forming double-stranded DNA (dsDNA) hairpin (HP) structures at each end. Based on the AAV genome, we constructed an AAV-ITR ssDNA minivector that consists of a GFP expression cassette flanked by both ITR sequences of 125 nucleotides. The minivectors were produced by digestion of the parental plasmids followed by denaturation. The self-complementary inverted T-shaped HP structure of the minivector was automatically formed. The HEK 293T cells were transfected with the AAV-ITR ssDNA minivector, plasmid, and dsDNA expression cassette. The results showed that AAV-ITR ssDNA minivector had relatively low gene expression efficiency in vitro. However, we found that the GFP expression efficiency of the D sequence-deleted AAV-ITR ssDNA minivector was significantly increased and was similar to those obtained with the plasmid and dsDNA expression cassette. Our data suggest that the AAV-ITR ssDNA minivector may be a new type of gene expression vector for gene therapy besides the virus and plasmid.
影响因子：
5.6
作者：
Xu-Shan Wang;S. Ponnazhagan;A. Srivastava
通讯作者：
Xu-Shan Wang;S. Ponnazhagan;A. Srivastava
影响因子：
12.4
作者：
McCarty, Douglas M.
通讯作者：
McCarty, Douglas M.