Fig. 4

Schematic diagram showing application of CRIPSR/Cas9 system in human cells. a Schematic diagram showing the construction of sgRNA library and functional screening steps. These sgRNAs were created to target 291 human genes, and the sgRNA library was designed by the assembly to the backbone of the virus. Through the lentiviral infection, the sgRNAs were delivered into HeLaoc-SC cells and those cells stably expressing sgRNAs were selected by FACS for green fluorescence. After toxin treatment, the resistant cells were selected for PCR and high-throughput sequencing analysis. b Schematic depicting the genome-editing process. Human iPSCs were edited using Cas9 complex and donor plasmid by electroporation. Details for genome-editing experiments are shown and the GFP+ cells were collected and used for imaging. c Diagram of the injection of Cas9 mRNA and sgRNA into human 3PN embryos to edit target genes, and 8–16 cell stage embryos were further collected to examine genome editing efficiency