4.1 Article

Efficient Editing of an Adenoviral Vector Genome with CRISPR/Cas9

Journal

INDIAN JOURNAL OF MICROBIOLOGY
Volume 61, Issue 1, Pages 91-95

Publisher

SPRINGER
DOI: 10.1007/s12088-020-00905-3

Keywords

Adenoviral vector; CRISPR-Cas9; T-cell therapy

Funding

  1. National Natural Science Foundation of China [81703053, 21771042]
  2. Guangdong Basic and Applied Basic Research Foundation [2018A030313860, 2020A1515010889, 2018A030313114]
  3. Guangzhou Science and Technology Project [202002030477]
  4. Innovative and Strong School Project of Guangdong Higher Education Institutions [2017KZDXM049, 2017KCXTD020]

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Immunotherapy using genetically modified T cells is crucial in treating tumors and viral infections, but efficiency of adenoviral vectors in infecting human T cells is a challenge. A simple and efficient method for editing adenoviral vectors using CRISPR-Cas9 to target the adenoviral fiber gene has been successfully established.
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 adenoviralfibergene.

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