Figure 7
Cellular chain reaction systems through sequential repair of loxP mutations. (A) Schematic representation of the chain reaction by repair of TATAloxP mutations to express EGFP in 293 T cells. (B) Experimental scheme used in (C). 293 T cells were transduced with the EGFP/different TATAloxP cassette with the indicated T > C mutations, then infected with Cre-R32V, and then transfected with nCas9-CDA and sgRNA targeting the T > C mutation. (C) Fluorescence images of 293 T cells expressing EGFP/TATAloxP cassette, Cre-R32V, and nCas9-CDA along with the indicated sgRNAs. When the EGFP/TATAloxP cassette was used with T > C mutations at the 11th and 13th bases in both arms, no EGFP expression was detected, even in the presence of the TATAloxP-targeting sgRNA (bottom right). (D) Schematic representation of barcoding method.
