Integrase-Deficient Lentiviral Vector as an All-in-One Platform for Highly Efficient CRISPR/Cas9-Mediated Gene Editing
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© 2017 The Author(s) The CRISPR/Cas9 systems have revolutionized the field of genome editing by providing unprecedented control over gene sequences and gene expression in many species, including humans. Lentiviral vectors (LVs) are one of the primary delivery platforms for the CRISPR/Cas9 system due to their ability to accommodate large DNA payloads and sustain robust expression in a wide range of dividing and non-dividing cells. However, long-term expression of LV-delivered Cas9/guide RNA may lead to undesirable off-target effects characterized by non-specific RNA-DNA interactions and off-target DNA cleavages. Integrase-deficient lentiviral vectors (IDLVs) present an attractive means for delivery of CRISPR/Cas9 components because: (1) they are capable of transducing a broad range of cells and tissues, (2) have superior packaging capacity compared to other vectors (e.g., adeno-associated viral vectors), and (3) they are expressed transiently and demonstrate very weak integration capability. In this manuscript, we aimed to establish IDLVs as a means for safe and efficient delivery of CRISPR/Cas9. To this end, we developed an all-in-one vector cassette with increased production efficacy and demonstrated that CRISPR/Cas9 delivered by the improved IDLV vectors can mediate rapid and robust gene editing in human embryonic kidney (HEK293T) cells and post-mitotic brain neurons in vivo, via transient expression and with higher gene-targeting specificity than the corresponding integrase-competent vectors.
Published Version (Please cite this version)10.1016/j.omtm.2017.04.002
Publication InfoOrtinski, PI; O'Donovan, B; Dong, X; & Kantor, Boris (2017). Integrase-Deficient Lentiviral Vector as an All-in-One Platform for Highly Efficient CRISPR/Cas9-Mediated Gene Editing. Molecular Therapy - Methods and Clinical Development, 5. pp. 153-164. 10.1016/j.omtm.2017.04.002. Retrieved from http://hdl.handle.net/10161/15931.
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Associate Research Professor of Neurobiology
Kantor Lab, the Duke University Viral Vector Core, facilitates the use of virus-mediated tools for gene transfer by investigators across diverse fields of study such as systems neuroscience, stem cell biology, metabolism, aging, cancer biology and others. Services are open to researchers within the Duke University Medical Center, Duke University, as well as outside investigators, worldwide.