Efficient Editing of an Adenoviral Vector Genome with CRISPR/Cas9
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Efficient Editing of an Adenoviral Vector Genome with CRISPR/ Cas9 Qiang Li1,2 • Hui Wang1,2 • Chen-yu Gong3 • Zhao Chen1,2 • Jia-xing Yang1,2 Hong-wei Shao1,2 • Wen-feng Zhang1,2
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Received: 3 February 2020 / Accepted: 26 August 2020 Ó Association of Microbiologists of India 2020
Abstract Immunotherapy based on genetic modification of T cells has played an important role in the treatment of tumors and viral infections. Moreover, adenoviral vectors engineered with improved safety due to their inability to integrate into the host genome have been key in the clinical application of T cell therapy. However, the commonly used adenoviral vector Ad5 exhibits low efficiency of infection of human T cells and the details of the intracellular trafficking pathway of adenoviral vectors in human primary T cells remains unclear. Resolution of these issues will depend on successful modification of the adenoviral vector. To this end, here we describe the successful establishment of a simple and efficient method for editing adenoviral vectors in vitro using the CRISPR-Cas9 gene editing system to target the adenoviral fiber gene. Keywords Adenoviral vector CRISPR-Cas9 T-cell therapy Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12088-020-00905-3) contains supplementary material, which is available to authorized users. & Hong-wei Shao [email protected] & Wen-feng Zhang [email protected] 1
Guang Dong Province Key Laboratory of Biotechnology Drug Candidates, Guang Dong Pharmaceutical University, Guangzhou, People’s Republic of China
2
School of Biosciences and Biopharmaceutics, Guang Dong Pharmaceutical University, Guangzhou, People’s Republic of China
3
Rundo International Pharmaceutical Research and Development Co., Ltd., Shanghai, People’s Republic of China
Two clinical reports on chimeric antigen receptor (CAR) modified T-cell therapy (CAR-T) published in 2018 cast doubt on the clinical use of lentiviral vectors [1, 2], indicating the urgent need to find efficient and safe delivery systems for use in CAR-T clinical therapy. A prevailing option is the use of adenoviral vectors with potentially increased safety due to the fact that they do not integrate into host cells [3]. Adenovirus vector serotype 5 (Ad5) has become the most commonly used adenoviral vector in gene therapy due to the clear genetic background [4]. However, because the natural cell surface receptor for this virus is the coxsackie receptor, which is expressed at low levels on T cells, the efficiency of Ad5 infection is low [5], limiting its use in the genetic modification of T cells. Adenovirus serotype 35 is a subgroup B adenovirus that recognizes CD46, which is highly expressed on human T cells, as its natural cellular receptor [6]. Meanwhile, the intracellular trafficking pathway of adenovirus in target cells influence the infection efficiency. However, current studies of the intracellular trafficking of adenovirus in target cells have focused on tumor cell lines, while the intrac
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