Non-viral gene delivery by minicircle vector expressing type VII collagen for the treatment of recessive dystrophic epidermolysis bullosa

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Title: Non-viral gene delivery by minicircle vector expressing type VII collagen for the treatment of recessive dystrophic epidermolysis bullosa
Authors: Qin, Yue
Advisor: Wang, Wenxin
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Date: 2017
Abstract: Recessive Dystrophic Epidermolysis Bullosa (RDEB) is a rare genetic skin disorder caused by mutations in the COL7A1 gene. The lack of type VII collagen results in absent anchoring fibrils which may cause the separation of the epidermis from the dermis in response to minimal trauma or friction. RDEB patients suffer from severe blistering and chronic wounds with their lifetimes. Unfortunately, there is no effective treatment for RDEB to date. Among all the currently investigating methods, gene therapy directly correct COL7A1 gene expression in the targeted cells. Compared to viral gene vector, the non-viral vector is a realistic alternative that increases gene cargo capability and decreases the safety risks. However, the bacterial backbone on the non-viral vector and the relatively large size of COL7A1 limits its transfection efficiency and the protein restoration. Here, an efficient non-viral minicircle vector was investigated by removing the bacterial backbone which includes prokaryote origin and antibiotic resistance gene. The reduced vector size can significantly increase the transfection efficiency. In addition, the silencing effect of the target gene can be decreased with lower immunoreactivity which leads to a long-term expression of type VII collagen. In this thesis, proper transfection approach was firstly selected from current commercially available reagents; then, the transfection efficiencies between minicircle and parental plasmid vectors were compared in different cell types. Minicircle vector with COL7A1 gene was constructed. Our preliminary results suggested that the minicircle-COL7A1 gene vector significantly increased the transfection efficiency and prolonged the expression of type VII collagen in vitro. To further improve the minicircle system, a tissue-specific promoter driving COL7A1 gene expression in Homo sapiens was introduced into the minicircle vector for future systemic delivery study. The minicircle system has shown promise as a potential RDEB gene therapy treatment.
Type of material: Master Thesis
Publisher: University College Dublin. School of Medicine  
Qualification Name: M.Sc.
Copyright (published version): 2017 the author
Keywords: COL7A1MinicircleNon-viral gene therapyRecessive Dystrophic Epidermolysis Bullosa (RDEB)Type VII collagen
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Language: en
Status of Item: Peer reviewed
Appears in Collections:Medicine Theses

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