Figure 6.

Viral vector methods for delivery of CRISPR. (A) AAV delivery of Cas9 and sgRNAs disrupting mutations in the Dmd gene in adult mdx mice, resulting in improvement of muscle biochemistry and function. Adapted with permission from Long et al. (2016). Copyright 2016 American Association for the Advancement of Science. (B) AAV intratracheal instillation delivery of sgRNAs in Cre-dependent Cas9 knock-in mice, resulting in lung adenocarcinoma (EGFP-positive tumors). Adapted with permission from Platt et al. (2016). Copyright 2014 Elsevier Inc. (C) A split Cas9 system in which the Cas9 C-terminal is packaged into one AAV vector and the Cas9 N-terminal is packaged into a second AAV vector. Reconstitution results in a fully functioning Cas9. Reprinted from Truong et al. (2015). Copyright 2014 The Authors (CC BY license). (D) AdV delivery of Cas9 and sgRNA targeting the Pten gene in mouse liver resulting in Pten mutation (see arrows by gel), and massive hepatomegaly and features of NASH in infected livers. Immunohistochemistry shows loss of Pten staining (arrows) one month after AdV infection; H&E stained micrographs show sections of steatosis (lipid accumulation, arrows) four months post infection. Adapted with permission from Wang et al., 2015. Copyright 2015 Mary Ann Liebert, Inc. Publishers. (E) Schematic of a lentivirus and CRISPR-based gene library functional screen used to identify the genes essential for West-Nile-virus-induced cell death. Reprinted from Ma et al. (2015). Copyright 2015 The Authors (CC BY license).