Figure 5.

Physical methods for delivery of CRISPR. (A) Microinjection disrupting two genes (Ppar-γ and Rag1) in Cynomolgus monkeys from a single injection into one-cell-stage embryos. Photographs of Founder Monkeys A and B, PCR products of the targeted loci from genomic DNA of A and B, and a control wild-type Cynomolgus monkey (Con). Adapted with permission from Nui et al. (2014). Copyright 2014 Elsevier Inc. (B) Electroporation delivery of CRISPR RNP targeting genes impacting mice coat color (Tyr) followed by transfer to pseudopregnant mothers. Bar plot quantifies coat color phenotypes generated from microinjection and electroporation at 1 ms pulse length and 3 ms pulse length. Adapted with permission from Chen et al. (2016). Copyright 2016 American Society for Biochemistry and Molecular Biology. (C) Hydrodynamic injection of CRISPR into mice results in liver-specific targeting (see bioluminescence image of hydrodynamically injected luciferase plasmid), generating indel mutation of two tumor suppressor genes and oncogenes. The development of liver tumors can be seen in the hematoxylin and eosin (H&E) and cytokeratin 19 (Ck19)-stained micrographs. Adapted with permission from Xue et al. (2014). Copyright 2014 Macmillan Publishers Ltd: Nature.